VCE Biology Unit 3&4
VCE Biology 3/4 Notes
Procedure - The set of processes used to define the scientific method typically including a hypothesis, experiment, observation, analysis, conclusion and evaluation
Hypothesis - If the (DV) then the (IV)
Independent variable - Variable that is manipulated and has a direct effect on the DV
Dependent variable - the measured variable that is observed/recorded
Repeatability - The ability of the experiment to be repeated identically in the same environment
Precision - The degree to which the results are consistent
Accuracy - The degree to which the results are close to the true value
Reproducibility - The ability of the experiment to be repeated with the same materials and method in a different environment
Validity - The experiment is measuring what it's supposed to be measuring
Bar Graphs - Different groups of data
Line graphs - Ranged set or continuous data
Control set up - The experiment without the independent variable
Controlled variables - Variables that are used to keep the experiment and control set up the same except for the independent variable.
Procedure - The set of processes used to define the scientific method typically including a hypothesis, experiment, observation, analysis, conclusion and evaluation
Hypothesis - If the (DV) then the (IV)
Experiment
Observation
Analysis
Conclusion
Method
Investigation
Independent variable - what is manipulated and has a direct effect on the DV
dependent variable - the measured variable that is observed/recorded
Experimental Set-up - To test the hypothesis, will have the independent variable
Control Set-up - Used to compare and confirm findings in experimental set-up, Does not have the IV,
Controlled Variable - Factors that are applied in both setups. Constant factor. purpose is to ensure that only one variable is tested at a time in an investigation: To make it valid. Eg. how much water you drink, weather, light, etc.
Repeatability
Precision
Accuracy
Reproducibility
Set of procedures to gain knowledge
6 key steps
Question
Hypothesis: educated guess or prediction, must be testable, states variables, “if, then”
Experiment
Observation
Analysis
Conclusion: statement of whether the original hypothesis was supported or refuted
Steps are not always linear but systematic
To make a hypothesis: ask a research question then change that into hypothesis
VCAA formula: If (DV) (relationship to IV) then (trend/effect on DV) and (IV whatever)
Controlled variable ≠ control set up
An investigation should only have ONE independent variable
Two setups only: Experimental Set-Up + Control Set-up
Results will be in graphs or tables.
RULES:
Title your table, including IV and DV
Column headings - Units included in Headings
Left column must be the IV
Data entry must have the same number of decimal places
Keep things neat and aligned
TAILS
Title
Axis
Interval
Label
Scale
Bar graphs are for different groups of data
Line graphs are a ranged set of data/ continuous data
Don’t forget repeatability
Example
Observation: Mice that are given Vitamin D increase the Calcium Absorption
Independent Variable: Vitamin D being given to mice
Dependent Variable: The Calcium absorption in the mice blood
Hypothesis: If Mice are given Vitamin D then the Calcium absorption in their blood will be increased.
OR
If mice are given Vitamin D then the Calcium levels in the blood will be less than the mice not given Vitamin D
Experimental set-up
100 Mice given Vitamin D
Vitamin D
Controlled Set up
100 Mice not given vitamin D but instead a placebo
Controlled Variables:
Amount of Vitamin D and placebo
Duration
Species of mice
Age of Mice
Same period of testing
Same food
Same water
Mice Sex
Results to support hypothesis:
A greater % of mice in the experimental group will have less calcium the blood while the control group show no increase of calcium absorption
Results to refute hypothesis:
A greater % of mice in the experimental group show no change in calcium levels
Accuracy
Not a quantity
How close to the true value
Bottle weighing 3kg measuring at 2kg is inaccurate
More trials does not increase accuracy
Precision
How closely two of more measurements agree with each other
Little spread among the values
If true value is 2.4 and results are 2.3, 2.3, 2.4, 2.4 then accurate and precise
5.6, 5.7, 5.9, 5.8 precise not accurate
Lack of accuracy but precision is systematic errors
Lack of precision could mean random errors
More trials can increase precision
Repeatability
Same method on identical materials with the same condition
The exact same experiment with the same person who did the experiment or operator
To obtain same results
Provides evidence of validity and reliability
Reproducibility:
Same method, same materials but different conditions
The exact same experiment in different environment with possibly a different operator
Identifying random or systematic errors
To obtain the same results
Validity
A measurement is valid if what is being measured is exactly what was claimed
Data is valid if the independent variable is the ONLY variable
Similar to accuracy
Reliability
Similar to precision
Random Errors
Random errors are unpredictable events or mistakes that affect the results
Outliers
Mistakes etc.
Affects accuracy and precision
Systematic Errors
Occurring the measuring system and affects every single measurement
Does not necessarily affect precision, affects accuracy
Re-calibrate machines
U3AOS2
Enzymes → P1
Photosynthesis → P2
Respiration → P3
Catalyst
Proteins
Enzymes
Enzyme functions
Catalyse
Substrates
Coenzymes
Active Site
Enzyme substrate complex
Enzyme product complex
Anabolic - building up, 1 + 1 → 2, always requiring energy, (endergonic)
Catabolic - breaking down, 2 → 1 + 1, reduce complexity, always releasing energy (exergonic)
Endergonic - Requires energy, uphill reaction, energy requiring
Exergonic - Releases energy, downhill reaction, energy release
Activation energy
Denature - Active site has changed and thus the enzyme becomes useless
Optimum temperature
Saturation point
Substrate concentration
Cofactors
Metal ions
Key Knowledge:
Enzymes and coenzymes are catalysts that assist with photosynthesis and cellular respiration
Factors that impact on enzyme function
Enzymes are proteins made from amino acids
Reusable
Speeding up chemical reactions by lowering the activation energy
Usually have the suffix ‘ase’ or ‘in’ (Eg. sudcrase, lipase, trypsin, pepsin)
Names can identify the substrate
Sucrase catalyses sucrose
Lipase catalyses lipids
Describes what the enzyme does or what fits in it
Substrates are substances that ‘fit’ into the active site of an enzyme.
Bonds are formed between the substrate, energy → water released and peptide
Active site is the region where only specific substrates are able to bind and undergo a chemical reaction
Substrate is the substance that can bind to the active site in an enzyme,
Enzyme specificity is based on the specific shape of the active site
When an enzyme binds to a substrate its called an enzyme substrate complex
Becomes enzyme product complex after catalysis
Theories:
Substrate and enzyme are directly complementary
Lock and Key Model
Perfect fits designed for each other
The active site and substrate are complementary
Induced Fit, the enzyme conforms for the substrate
The active site becomes complementary
Enzymes ARE proteins and are made of amino acids
Biological catalysts → speeds up processes
Not permanently changed in processes
Reusable
NOT reactants → written on the arrow in reaction equations
Catabolic Reactions
Large molecules to small molecules
Breaks down
Exergonic, releases energy
Needs water
Anabolic Reactions
Small molecules to large molecules
Builds
Requires intake of energy
Endergonic
Produces water
Activation energy:
Energy needed for any chemical reaction, any reaction will occur with enough energy. Enzymes work by lowering the required activation energy.
Factors affecting enzyme activity:
Temperature
Heat energy means more collisions between enzymes and substrates however enzymes denature so the rate of reaction falls
Optimum temperature is 37.5 degrees celsius
When there is low temperatures there is less kinetic energy
The enzyme is NOT denatured at low temperatures but it IS denatured at high temperatures
Low kinetic energy low molecule collisions less enzymes substrate complexes forming etc.
pH
pHs have an optimum pH for enzymes but enzymes can denature on both sides of pH, if it’s too low or too high.
Works only within a small pH range
pH disruption generally results in complete loss of activity
Identical parabola
Substrate concentration
Substrate graph optimum is the point of saturation
Point of saturation means all active sites have been occupied and the enzymes must produce at a stable reaction rate.
More enzymes = Increasing reaction rate
Increasing then plateau
Enzyme concentration
Rate of reaction can increase as long as there is enough substrate
If there is an overabundance of enzymes and not enough substrate no reaction will occur
Constant line
Cofactors & Coenzymes
Cofactors can either be metal ions or coenzymes
Metal ions bridge enzymes + substrate together, combined with the catalyst
Coenzymes, non-proteins, organic complex
makes the substrate fit better
All coenzymes are cofactors not all cofactors are coenzymes
Activate the enzyme
Inhibitors
Competitive and noncompetitive inhibitor
Competitive is COMPETING with the active site and binding to the active site
Non competitive inhibitors are not trying to bind to the active site, they bind to the enzyme changing the active site inhibiting the substrate from binding
More substrates can overcome competitive inhibition but cannot overcome noncompetitive inhibitors
Competitive Inhibitors are TEMPORARY and REVERSIBLE, because they can eventually leave the enzyme
Allosteric = not the active site
Non competitive inhibitors force the enzyme to change PERMANENTLY and are IRREVERSIBLE
Stirring/Agitation
Increases collision and increases reaction rate
Increases the substrate and enzyme collision
Feedback inhibition
Metabolic pathway
Dephosphorylation
Phosphorylation
Large molecules need to be broken down slowly over multiple stages
The last product made in a process can be an inhibitor for enzyme one which stops too much product being made.
Feedback inhibition
Metabolic pathway
Each pathway requires a specific enzyme
The pathway is stopped by feedback inhibition
If the gene coding for an enzyme is messed up then the lack of feedback inhibition will cause a build up of a certain substrate and there will be no product
Exergonic
Dephosphorylation - Losing a phosphate
Phosphorylation - gaining a phosphate
ATP
ADP
Phosphate
Metabolic Reactions
Mechanic work
Nucleotide
Hydrolysis
Hydrolyse
Loaded - Fully energised (ATP)
Unloaded - Not fully energised (ADP)
Inorganic phosphate (Pi)
Organic - molecule with carbon
Inorganic - molecule without carbon
Cells need energy to make muscles work, carry out chemical reactions, the growth and repair of cells, making larger molecules, maintaining body temperature
Enzymes are important in cellular respiration
Mechanical reactions, metabolic reactions
Adenine Triphosphate
1 Adenine (N-Base) + 1 Ribose Sugar + 3 Phosphates
Linked by hydrogen bonds
ATP breaks down and releases a lot of energy and becomes ADP and phosphate
ATP → ADP → ATP (reversible)
ATP is a nucleotide
Hydrolysis - To split ATP
Water is needed to split ATP into ADP
ATP is a loaded molecule
Enzyme used to catalyse ATP is called ATPase
Reaction type is Exergonic, Hydrolysis, Catabolic, Dephosphorylation
ADP is an unloaded molecule
ATP can act as a coenzyme and assist other enzymes
It is an energy carrying molecule
Provides enough energy to support reactions through the breaking of phosphate bonds.
ATP’s third phosphate is weakly bonded and has high energy
ATP is synthesised through cellular respiration
ATP synthase is enzyme to create ATP from ADP
Located in the mitochondria’s membrane and in chloroplasts
Found where ATP is needed to be made ie: mitochondria, chloroplast etc.
Dehydration/Condensation reaction = water being produced (ADP→ATP)
Endergonic, Anabolic
OTHER COENZYMEs:
Coenzymes are not specifically for substrates they are carriers to the reaction products
Coenzymes are regenerated to be reused
H+ (electrons)
NAD+ (+) H+ (+) 2e- (Unloaded → NADH (Reduction) (Loaded)
The other way round is Oxidation
FAD (Unloaded) → FADH2 (Loaded)
NADP→ NADPH
Workbook 20-21
CELLULAR RESPIRATION
Reactants
Cellular resp
Glycolysis
Krebs cycle - Takes place in the mitochondrial matrix
ATP yield (NOT NUMBER IN TEXTBOOK)
Electron transport chain - Set of reactions in the mitochondria
inputs/outputs
Locations of glycolysis
Coenzymes
Pyruvate/Pyruvic Acid (3 Carbon molecule, break down of glucose)
Intermediate reaction
Cristae - Mitochondrial inner membrane, most ATP is made
Mitochondrial matrix is the fluid in the mitochondria
Glucose + Oxygen ⇒ 30-32 ATP + (Water) + (Carbon Dioxide)
CO2 and Water are byproducts the intention is for ATP
1 glucose makes 30-32 Total ATP which is too much energy to be produced at once
Glucose | Oxygen | Carbon Dioxide | Water | ATP | |
Animal | Ingested | Inhaled | Exhaled | Output | Energy currency to maintain life |
Plants | PHS product | PHS product | PHS input | PHS input |
All cells respire
Respiration is a set of metabolic reactions
Purpose: To convert glucose to ATP
Mitochondria is the powerhouse of the cell
Wherever ATP is being made there is ATPsynthase
When glucose enters a cell it breaks down with glycolysis, taking place in the cytoplasm
The glucose breaks into Pyruvate x 2which breaks down into CO2 x 2 and Acetyl CoA x 2
Pyruvate to CO2 and Acetyl CoA is an intermediate reaction
Acetyl CoA then goes to the mitochondria’s matrix which is a fluid where the Krebs cycle occurs
When glucose → pyruvate → acetyl CoA is made Hydrogen ions are released which would disrupt the pH of the cell, NAD + FAD pick up the hydrogen ions and become NADH or FADH2 which are reduced, loaded molecules and are used to transport to the cristae
NAD and FAD are electron carriers/coenzymes
NAD is an important coenzyme that is used to activate lactic dehydrogenase enzyme
Intermediate reaction = link reaction = transition reaction
Glycolysis → Intermediate reaction → Krebs cycle → Chemiosmosis
Glycolysis produces 2 ATP and Krebs produces 2 ATP and Electron transport chain produces 26-28 ATP so in TOTAL 30-32
Glycolysis - Cytosol
Matrix - Kreb cycle
Cristae - ETC
Mitochondria has;
Two membranes, Outer and Inner (Cristae)
Shaped for maximum efficiency
Requires 2ATP and ATPase
Breaks down into Pyruvate or Pyruvic acid
NAD+ come and collect the hydrogen ions
2 ATP is produced from both ‘breakdowns’
Total of 4 ATP produced but net 2ATP because there is an investment of ATP
Input 2 acetyl coa 2 Adp 6 NAD+ 2 FAD+
Output 4CO2 2 ATP 6 NADH 2 FADH2
Only 2ATP are produced
Oxygen is NOT involved
More electrons are picked up by NAD and FAD
All co2s have been released
Made in the MATRIX
NADH is oxidised and GIVE UP its hydrogen ions
Made in the Inner mitochondrial membrane or the cristae
The hydrogen ions cross the cristae membrane
All the hydrogen ions then go through ATP synthase and make ADP into ATP
26-28 ATP
Facilitated diffusion through ATP synthase
Electron transport chain = oxidation phosphorylation
H+ cross membrane → proton gradient (concentration) increases outside membrane → H+ is then facilitated diffusion-ed through ATP synthase → ATP made
Oxygen is the final electron acceptor, water is made as oxygen + H+ + electrons
10 mill ATP is produced per second by one cell
Fermentation
Alcohol Fermentation
Lactic Acid Fermentation
Anaerobic Respiration
Muscle Fatigue
O2 Debt
Ethanol
Lactic Acid
When oxygen is not present in the cells, the process of fermentation occurs
Glycolysis → Fermentation
NADH returns the H ion and the pyruvate becomes lactate or ethanol
Reversible once oxygen becomes available
Lactic acid in animals, bacteria and some fungal cells
Alcohol/Ethanol produced in plants and yeast
2(C3H6O3)
Lactic acid is poisonous to the body and builds up creating muscle cramps, it is broken up by oxygen
Ethanol is 2C2H5OH created by plant and yeast fermentation
Lactate dehydrogenase and Alcohol dehydrogenase
Glucose → ethanol + carbon dioxide + energy (2ATP
Glucose → Lactic acid + energy (2ATP)
When animals run out of O2, pyruvate becomes lactic acid creating muscle fatigue and O2 debt
Provides rapid bursts of ATP in muscle cells
But is incredibly toxic
Oxygen concentration
Temperature
Glucose availability
Hydration, light, age, activity level
Respirometer is a device that determines an organisms respiration rate by measuring the rate of exchange of O2 and CO2
Living specimens enclosed in sealed container
Pressure changes affect the manometer
Increasing Co2 levels or decreasing oxygen levels → Increases respiratory rate
**RATE OVER TIME (over time must be included)
Temperature denatures enzymes therefore pathways cannot continue
Oxygen measured by the amount of CO2 produced
Increase O2 —> increase resp. Rate until point of saturation
Anaerobic respiration is measured by lactic acid of alcohol produced, aerobic respiration measured by Co2 levels
Glucose and oxygen are substrates
Glucose conc increases
Biofuels - reducing the amount of greenhouse gases by being renewable and recyclable fuel made from recently living organisms such as plants and algae
Fossil Fuel
Biomass
Fermentation
Renewable
Burning biofuels releases carbon dioxide but they are carbon neutral because they cancel out the amount they release by holding that amount of carbon dioxide when being grown.
Needs to be easy to make, transport and able to mix with fossil fuels
Places to grow this.
Processes requiring energy
Feedstock | Advantage | Disadvantage |
Corn, Rapeseed |
|
|
Waste cooking oil, Lignocellulosic feedstock |
|
|
Microalgae |
| Low lipid content Contamination problem |
Engineered microalgae |
|
|
Two main types of biofuels, bioethanol and biodiesel
Bioethanol/Ethanol
Fermenting sugarcane /starchy plant materials
Blended with petrol
Biodiesel
Made from lipids/fatty acids
Inputs, outputs
Roles of enzymes and coenzymes in facilitation
Locations of light dependent and light independent stages
Rate of photosynthesis and factors that affect rate
Factors: light, water, temperature, carbon dioxide
Photosynthesis
Light dependent
Light independent
Autotrophs, producers
Heterotrophs
Palisade layer - generally the more photosynthetic cells
Epidermal layer
Spongy layer
Mesophyll - all the in between bits (palisade and spongey)
Bundle Sheath cells
Stomata/Stomate
Guard Cells around the stomata - close at night, open day which allows for the diffusion of Co2
Oxidation
Reduction
Xylem tubes - Carry water
Phloem tubes - Carry food
Veins
Granum is the pile of discs or Thylakoids
Thylakoid, the membrane of the discs in a chloroplast, is the disc
Stroma
Inner Membrane
Outer Membrane
NADP and NADPH
Calvin cycle
Limiting Factor
G3P
PG3 - Phosphoglycerate
Glucose
C3
C4
CAM
Turgid
Flaccid
Sunlight is needed because it is the initial energy source
Plants make glucose during daylight, as long as there is sunlight
They require glucose because they are autotrophs, they make their own food
Water is diffused by osmosis
Inputs: Water + Carbon Dioxide
Outputs: Sugar + Oxygen
Input/Output and Light | Location | How |
Carbon Dioxide | Stomata | Gas |
Water | Roots | Osmosis |
Light + Chlorophyll | Leaves, Chlorophyll | Chlorophyll which traps light |
Sugar/Glucose | Stored as Starch and used in mitochondria in leaves and phloem | |
Oxygen |
Include light and chlorophyll on the →
Cuticle protects plant
Epidermal Layer is made of epidermal cells
Palisade Layer, made of palisade cells
Sponge layer, made of sponge cells and air
Stomata used for co2 diffusion and water transpired out
Photosynthesis takes place in the chloroplast
Chlorophyll, containing ATP synthase
6CO2 + 6H2O → C6H12O6 + O2
6CO2 + 12H2O→ C6H12O6 + 6O2 + 6H2O
Reduction is gain of electrons
Oxidation is loss of electrons
H2O is oxidised, CO2 is reduced
NADP comes and picks up the hydrogen ions released from the reaction and split of H2O and comes and picks up the ions and becomes NADPH
Oxidation reaction = water being oxidised to oxygen through NADP
Reduction reaction = Carbon Dioxide is reduced to glucose
Glucose is made for cellular respiration
Xylem tubes carry water from roots to leaves and parts of the plant through veins
Phloem tubes carry food to all parts of the plant body for all cells to respire
Made up of granums which are discs filled with chlorophyll and membrane known as Thylakoids
Water is trapped in the Thylakoids and is split releasing Oxygen to be diffused out of the cell, some may be used in respiration but the rest will diffuse into the atmosphere
NADP comes and collects the hydrogen from the split H20 becoming NADPH
The H+ is used to synthesise glucose
Light dependent stage, splitting of H20 takes place in the Thylakoid
The Calvin Cyle or the synthesis of glucose and the reduction of Co2 doesn’t require light and is the light independent stage
ADP + Pi becomes 18ATP
Photosynthesis is photo, then synthesis
Photo - light dependent
Thylakoid/Granum
Chlorophyll traps the light to split water and produces 18 ATP and O2
Synthesis - light independent
Stroma
Calvin cycle
Using 18 ATP from light dependent reaction
Photosystems 1 and 2 are photosynthetic pigments that absorb light energy
Calvin cycle: CO2 + NADPH + 18ATP → NADP + 18ADP + GLUCOSE
Occurs in the stroma and doesn’t require light but depends on products of light dependent stage
Uses ATP and NADPH to synthesise glucose
Endergonic and Anabolic
NADPH → NADP
Enzyme rubisco is required
It fixes RUBP (sugar with 5 Carbon) to CO2 and turns it into a 6 Carbon molecule
Which becomes 2 x G3P
One G3P will leave as an output of SUGAR
The other will continue in the cycle and become RUBP
A higher concentration of CO2 means more reaction
Calvin cycle runs 6 times before producing glucose
Rubisco is an enzyme for Carbon Dioxide but when there is a high concentration of Oxygen, oxygen will bind to Rubisco instead.
External Factors include:
Light
Temperature
CO2
Water
Internal factors
Chlorophyll - affects the amount of light being absorbed - is a limiting factor
Limiting factors are factors that limit the reaction such as limited by temperature being too low or temperature being too high etc.
Photorespiration
When there is no CO2 entering due to closed stomata and increase in O2 concentration then photorespiration occurs
Rubisco picks up O2, high temp, stomata closed, when oxygen is more concentrated than co2 and creates CO2 by binding O2 to Rubisco
Stomata/Stomate - openings guarded by the guard cells
Guard cells turgid/Stoma Open
Turgid = Full
Cells flaccid/Stoma closed
Guard cells use osmosis to open and close by the concentration of sugars
C3 plants have thin bundle sheath cells
C3 make up most plants
Occurs only in mesophyll cells
C4
Pep Carboxylase ( PEPCase) always binds to CO2 to produce C4 Acid Oxaloacetate
More efficient than Rubisco because it will always collect Co2
PepCase makes Malate and CO2 in the Mesophyll cells from Co2 and PEP Carboxylase
Co2 Increases, Rubisco picks up Co2 to fix to RUBP and continue the calvin Cycle
Occurs in Bundle sheath then Mesophyll
C4 | C3 |
2 Carbon Fixations | 1 Carbon Fixation |
Photosynthesis | Photorespiration and Photosynthesis |
Adapted to low light and low water, more efficient | Not as good in hot weather, closes stomata in hot weather |
Thick Bundle Sheath Cell | Thin bundle sheath cell |
1st product is Oxaloacetate (4p) | 1st product is PGA (3p) |
CAM
Closes stomata during the hottest part of the day
Traps Co2 at night time
Making sugar with trapped CO2
Similar to C4 plants uses PEP carboxylase to malate fixation of CO2 and then to the Calvin Cycle
Occurs in Mesophyll all the time however the different processes are at night vs day
Comparing | C3 | C4 | CAM |
# of CO2 Fixation reactions | 1 | 2 | 2 |
1st stable product from Co2 Fixation | PGA (3C) | Oxaloacetate (4C) (Malate) | Malate (4C) |
Where Calvin Cycle occurs | Mesophyll cells | Bundle Sheath Cells | Vacuole & Mesophyll cells |
Photosynthesis and/or Photorespiration | Photosynthesis and Photorespiration | Photosynthesis | Photosynthesis |
Efficiency of CO2 Fixing | Poor | Good | Good |
When Stomata open | Day | Day | Night |
Best adaptation | Moderate cool and wet | Hot and sunny | Very hot and Dry |
Proteins
Hormones
Structural proteins
Biomacromolecules
Carbohydrates
Lipids
Nucleic Acid
Monomers
Polymers
Polymerisation - Making polymers from monomers, a reaction that combines monomers to make polymers
Oligomers
Alpha helix
Beta Pleated sheet
Polypeptide
Peptide bond
Primary, secondary, tertiary, quaternary protein structures
Globular Protein
Filament
Protofilament
Fibrous Protein
Proteomics - Study of proteome
Proteome - the complete array of proteins produced by a single cell/organism in a particular environment is called the proteome of the cell/organism
Purines - A & G
Pyrimidines - T & C
Chromatins
Messenger RNA
Transport RNA
Ribosomal RNA
Transcription - Occurs in nucleus, copying Dna code onto mRNA
Translation - Translating mRNA (Decoding)
Proteins are used for everything such as contraction, reception, hormones, protection, transport, storage, enzymes, structural, identification, signal etc.
They are coded from Amino acids which are building blocks
Contain C, O, H, N & S
Proteins are building blocks. They have a wide range of functions
Monomers are the basic building block
Amino acids are the monomers for proteins
Nucleotides are monomers for nucleic acids
Monosaccharides are monomers for carbohydrates
All amino acids have
An amino group
A carboxyl group 0=C-OH
A unique side chain (often depicted as R) H-C-R
Amino Acid 1 + Amino Acid 2 → Dipeptide + Water
Condensation/Dehydration reaction because production/loss of water
Amino acids combine and peptide bonds form from amino acids to become a polypeptide.
The covalent bond between the amino acids is the peptide bond
Polypeptide chains can be broken down via hydrolysis reactions which splits the chain.
4 levels of protein structure
Primary protein structure - Linear sequence of a chain of amino acids
Secondary protein structure - an alpha helix + a beta-pleated sheet which is the folding of polypeptide chains into helices or sheets
Tertiary protein structure - 3D folding pattern of a protein due to side chain interactions - Globular protein, very specific shape being formed
Some proteins only continue to Tertiary structure but some combine into more than one amino acid chain
Quaternary Protein structure - Protein consisting of more than one amino acid chain
Linear sequence will provide information on how the protein folds, function or no function, evolutionary relatedness between species.
Hydrogen bonds hold alpha helix, peptide bonds form between amino acids
SECONDARY PROTEIN STRUCTURE
Alpha helix
Beta Pleated sheets
Random Coils
TERTIARY STRUCTURE
Total irregular folding and bending of chain
Causes amino acids to become close
Function depends on shape
3D protein
Disulphide bonds only existing in tertiary structure
Hydrogen bonds
QUATERNARY
Fibrous or globular
Some are conjugate containing inorganic compounds
Eg. Haemoglobin
The two types of nucleic acids
DNA + RNA
Made of a sugar, phosphate and nitrogenous base (ATGC or AGCU)
RNA is single stranded
DNA is double stranded and also has no oxygen - Antiparallel double stranded helix
AT are double bonded
GC are triple bonded
The phosphate makes DNA negatively charged
The units of DNA inside the nucleus are Chromatins which when needed to split become Chromosomes.
5’ and 3’ Antiparallel structure (‘ = prime)
Three types of RNA
Messenger RNA - Carries instructions for polypeptide synthesis
Ribosome - Structural subunits of the ribosome
Transfer RNA - Carries amino acids to ribosome
DNA are the instructions the RNA are the messengers
RNA copies a strand of DNA and then this is used to go to the ribosome which codes 3 bases at a time using amino acids taken from food and the breakdown of proteins
DNA is in the mitochondria, chloroplast and some in the mitochondria only 1 type
RNA in the nucleus and cytoplasm has at least 3 types
Made in 3s in the ribosome
Transcription occurs in the nucleus - the copying of DNA onto mRNA
Translation in the ribosome - decoding of mRNA into the ribosome
Gene Expression = Protein Synthesis
Transcription
Translation
Ribosome
Protein Synthesis
Sense/Antisense strand
Template/Non-template strand
Amino acids
Non-Coding/Coding
Pre-RNA
Elongation
Binding
Initiation
Promoter region
Methylated Capping
Introns
Exons
Gene sequence
Exon juggling
DNA codes the instruction for protein synthesis but
Ribosome is the site of protein synthesis which is in the endoplasmic reticulum
DNA cannot leave nucleus but Ribosomes cannot enter the nucleus
A copy of DNA is made as RNA through transcription
At the ribosomes, the copy is translated by tRNA and the necessary amino acids will be produced
Template strand/Antisense Strand/Non-coding strand
Sense/Non-template/Coding strand
Gene sequence = correct order of nucelotides
Steps of Transcription
1.Initiation: Initiation factors (proteins) bind to DNA stand to switch on the gene
2. Binding: RNA polymerase binds to the promoter region of the template strand
3. Elongation: RNA polymerase moves along the template stand, preliminary to RNA
Methylated Capping and poly-deniylation tail for Transcription
Introns and Exons
Base pairs are read by tRNA three base pairs at a time
tRNA structure consists of a structure of amino acid and anti-codon
TRANSLATION STEPS
mRNA attaches to a ribosome
tRNA (anticodon) attaches to mRNA (codon) (base pairing)
A specific amino acid is detatched to form either a polypetide or a peptide bond with an amino acid
Stop Codons are codons that literally stop the process of forming the polypeptide chain etc.
The stop codons are UAA, UAG, UGA
All living organisms have the same amino acids
There are more than one codon for each amino acid to allow for error
Gene Regulation - when a gene is switched on
Structural gene - gene that encodes for a specific protein
Regulatory gene - A gene that encodes for protein that regulates the structural gene ie activators or repressors to switch the genes on and off, codes for regulatory proteins
Activator proteins - turns genes on to start transcription
Repressor proteins - turns the gene off to prevent transcription
Promoter region - where RNA polymerase binds
Operator region - where regulatory proteins binds
Operons - only occurring in prokaryotic cells, a functional unit of transcription that regulates gene expression in bacteria
TRP Operon
Tryptophan
Anti-termination loops
Termination Loop/Attenuator stem loop
Hairpin loops
Attenuation
Attenuator
Lead mRNA
trpL
Leader region
Genes need to be regulated because not all cells can make every single protein all the time so certain cells are made to produce certain proteins and certain genes are on or off
The controlling of gene expression
Structural gene
Regulatory gene
The regulatory protein (activators or repressors) binds to the Operator region
Prokaryotic Gene Regulation
Does not have introns or exons or gene capping
Can combine multiple structural genes to create an operon
Occurs in the cytoplasm
TRP OPERON
The TRP operon is an operon in bacteria
Made by bacteria by the TRP Operon
Tryptophan - Amino Acid
The TRP operon codes for enzymes that catalyse the creation of Tryptophan
When TRP is high - Operon turns on
When TRP is low - Operon turns off
The TRP can regulate the levels
Only in prokaryotes
An Operon is a cluster of genes under the control of a single promoter
Transcribed and then Translated
When the operon needs to lower the amount of trp enzymes the regulatory region will make regulatory proteins that switch structural genes on and off
Tryptophan levels high → Tryptophan binds to the Repressor, and RNA polymerase cannot bind
ATTENUATION
Alternative method of reducing the expression of the trp Operon in prokaryotic cells
It relied on the capacity for prokaryotes to be both transcribing and translating SIMULTANEOUSLY
It prevents transcription from being completed
It can occur when tryptophan levels are high as a backup to terminate transcription when the repressor detaches from the operator
Stops further synthesis of the creation of tryptophan
In the trpL (lead protein)
The leader region of the operon is the trpL leader + Attenuator
trpL codes for the leader mRNA
Four regions that can form base pairs to form 3 hairpin loops
When there are low levels of tryptophan, the antiterminator loop occurs and blocks the formation of the termination loop (attenuator) it pauses the protein synthesis
The attenuator is like a stop codon and will stop transcription, the termination loop will cause the ribosome to stop translation
there are trp codons on the trpL when the tryptophan levels are high and it will pause the ribosome at the stop codons which covers the 2 and stops 2 and 3 from binding and then allowing the termination loop
When the 3rd loop forms stuff don't work
Vesicles - membrane bound packages
Transport Vesicles
Secretion Vesicles
Synthesis
Exocytosis - leaving the cell
Lysosomes - vesicles with digestive enzymes
Golgi apparatus
Smooth ER
Rough ER
Proteins are transported in vesicles from the Rough ER to the golgi apparatus and are then transported to be secreted from the cell membrane or used around the cell
Smooth ER for hormones and lipids
Rough ER usually for proteins
REVISION QUESTIONS:
Tryp Operon
What is the role of the trp operon in regulating the synthesis of tryptophan?
The TRP operon regulates the production of the enzymes for the biosynthesis of tryptophan.
What type of gene regulation is involved in the regulation of the trp operon?
Negative regulation.
How does the presence of tryptophan affect the expression of the trp operon?
HIgh levels of TRP make the trp repressor fall off the operator region by binding to the repressor, in low levels, the repressor falls off. If the repressor falls off while trp levels are high then attenuation is used.
What is the role of the trp repressor in regulating the trp operon?
It prevents RNA polymerase from transcribing the leader genes and the structure genes so that the enzymes for the biosynthesis of tryptophan cannot occur.
How does attenuation regulate the expression of the trp operon?
It acts as an emergency in case the repressor is not bound to the operator region of the operon when tryptophan levels are high. By not pausing RNA polymerase when it reaches region 1 and pausing at region 2 it allows regions 3 and 4 to bind creating a hairpin loop that forces the ribosome to fall off.
Gene Regulation
What is gene regulation and why is it important for an organism?
Gene regulation is the control of the production of proteins based on gene expression and it is important for specialised cell functions and proper cellular development.
What are the different types of gene regulation mechanisms?
Negative and positive regulation, splicing, attenuation, feedback inhibition, transcriptional regulation, translational regulation
What is the role of transcription factors in gene regulation?
How do epigenetic modifications affect gene expression?
What is the difference between positive and negative gene regulation?
Protein Packaging
What is protein packaging and why is it important?
What is the structure of a nucleosome and how does it package DNA?
How are histones involved in protein packaging?
What is chromatin and how does it affect gene expression?
How does the packaging of DNA affect the accessibility of genes for transcription?
Protein Synthesis
What is protein synthesis and what are the two main stages involved in it?
What is the role of messenger RNA (mRNA) in protein synthesis?
What is the function of ribosomes in protein synthesis?
How is the genetic code translated into a specific amino acid sequence?
What is the role of transfer RNA (tRNA) in protein synthesis?
Nucleic Acids
What are nucleic acids and what is their function in cells?
What is the structure of a nucleotide and how are nucleotides joined together to form a nucleic acid?
What is the difference between DNA and RNA?
What is the function of DNA in cells?
What is the central dogma of molecular biology and how do nucleic acids play a role in it?
Proteins
What are proteins and what is their function in cells?
What is the structure of an amino acid and how are amino acids joined together to form a protein?
What is the difference between a primary, secondary, tertiary, and quaternary structure of a protein?
What is denaturation and how does it affect the function of a protein?
What is the role of chaperone proteins in protein folding and quality control?
Biotechnology - use of an organism or organism component to make a product or process (ie: COVID vaccine)
DNA technology - sequencing, analysis and cut & paste of DNA
Meiosis
Genetic Recombination - Exchange of info between 2 DNA segments
rDNA - recombinant DNA technology
Vectors
Plasmid
Cloning Factor
Amplified Gene
Amplified Protein
Recognition sequence - identifies if something needs to be cut
Restriction enzymes
Reverse transcriptase
Ligase
Polymerase
Sticky ends
Blunt ends
Ligation
VCAA Knowledge: The use of enzymes to manipulate DNA including polymerase to synthesise DNA
Majority of Biotechnology relies on DNA manipulation
DNA technology is the sequencing, analysis and cut & paste of DNA sequences
Genetic Recombination which uses or occurs in meiosis and exchanges info between 2 DNA segments (like homologous chromosomes)
Occurs b/w same species
Makes a recombinant chromatid (or non-recombinant)
DNA manipulation literally manipulates genomes/genes to introduce or take away specific objectives.
rDNA is recombinant DNA technology
Genetic engineering, recombinant biotechnology, DNA manipulation
Gene technology includes manipulation and analysis of DNA
DNA manipulation alters DNA by adding or editing DNA
Bacteria is often used as vectors
To reproduce an edited gene a cloning vector is needed such as a plasmid from Bacteria to create a recombinant plasmid
Protein or Gene can be amplified
In order to → Tools used
cut DNA: Restriction enzymes (Endonuclease)
Stick DNA fragments: DNA ligase
Copy of DNA: Polymerase
Make multiple copies of DNA: Polymerase Chain reaction (PCR)
Separate DNA fragments: Electrophoresis
Edit Genes: CRISPR
To synthesise DNA, replicate or repair
Copy DNA and make copies
Example: Taq polymerase used in PCR
DNA Polymerase creates DNA by asembling 749 (approx) nucelotides per second
Synthesis from 5’ to 3’
Taq Polymerase is a DNA polymerase used on PCR (chain reaction)
Joins different pieces of DNA
Used to make recombinant DNA
Recombines the DNA
Ligase closes the ‘nicks’ in the phosphodiester bonds + close the gaps and seals the newly transferred dna segment
Same Restriction enzymes cut gene of interest and plasmid /cloning vector
Plasmid and gene fragments connect and anneal
Ligase used to anneal the recombinant plasmid and seal it
Ligation
Cuts DNA and creates fragments
Examples: BamH1 (Bam = bacteria, H = Strain, 1 = Order of discovery)
EcoR1 and Taq1
EcoR1 - GAATTC (recognition sequence) will cut between the G and A so long as there is an AATTC following.
Sticky ends and blunt ends when one is overhanging or when there is a clean cut
Obtaining DNA from RNA
Reversing the transcription
PCR
Heating and Cooling Cycles
Amplify
DNA segment
Target DNA sequence
Primers - complementary segments that will attach to the 3’ prime end of a chain, extend the DNA through complementary base pairing
Buffer Mix - Maintains pH
Taq polymerase -
PCR is a technique to amplify or produce copies of a DNA segment
Using heating and cooling cycles
Replication at an exponential Rate
3 Steps
Denature
Separating the hydrogen bonds (94-95o)
Anneal
Primers bind to template (50-56o)
‘Annealing of the primer when it ‘cools’
Extension
Increases in temperature to 72
Synthesising a new strand
PCR can be used for;
consumer genomics
Food and agriculture
Forensic science
Genetic research
Medicine
Phylogenetics
Environmental biology
State purpose of PCR
To replicate specific DNA segments
Identify components
Primer
Nucelotide
Heat
Buffer
DNA sample
PCR Tube
Draw Steps to illustrate basic process
Heat stuff up and break hydrogen bonds
Anneal stuff with primers at the 3’ ends
Synthesise new strand
Identify Two applications
Gel Electrophoresis - used for sorting dna fragments
Agarose Gel - Jelly (containing buffer solution to maintain pH)
Buffer
Gel Matrix
Cathode
Standard - Kind of like a ruler used to compare dna sizes
Loading Dye
Ethidium Bromide - Fluorescent dye, binds to dna, mutagen
DNA fragments put into gel, and has positive and negative charges on each side, DNA is negative charged cos Phosphates so is drawn to positive charge.
Gel/matrix traps larger fragments and therefore separates dna fragments by size
Separate mixtures
Calculate size
Components
Electric current
Agarose Gel - traps dna allowing small fragments to move further down
Loading Dye
Standar - to calculate sizes
Ethidium Bromide - Fluorescent dye, binds to DNA
Buffer solution
Combs - to create wells to load DNA
Positive and negative terminal
STEPS
DNA fragments loaded in neg side
Electric current
Dna fragments migrate towards pos side
Small fragments move easily and travel far distance/larger fragments are trapped and travel less
Unit, Size (bp or Base Pairs) , Quantity (ng)
Sometimes known as a ladder (the standard)
DNA profile
Polymorphic regions - regions that have more than one change
Polymorphisms - differences in these region
Probable Origin
Microsatellites - they are STRS
STRs/Short tandem repeats
Non-coding DNA
Repeating code
Genetic loci (location)
Heterozygous genotype
Combined DNA index System or CODIS
Can be used to identify probable origin
Reveal family relationships
Identify Victims in disasters
Paternity Tests
Find Evolutionary relationships between species
Small sections in DNA vary, everything else is identical, these small sections identify individuals
Everyone inherits unique combination of polymorphisms
Process: Collecting DNA then cutting with endonuclease and then using gel electrophoresis to separate and identify.
STRS are regions of non coding DNA that contain repeats of the same nucleotide sequence
Short → 1-9 base pairs long
Tandem → Repeating Code
STRS are found in different places/genetic loci
Microsatellites (1-9bp), Minisatellites (10-100bp), Macrosatellites (>100bp)
Minisatellites for fingerprinting
STRS/Microsatellites for DNA profile
STR-XX (x,y)
If it’s different length, then its heterozygous
If same length, homozygous
ETHIC ISSUES
Limits of testing
Advantages | Disadvantages |
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Plasmids
Bacterial Chromosome
Recombinant
Colony
Vector
Antibiotic plate
Transgenic organisms (TGO)
Genetically modified organisms (GMO)
Cloning Vector
Gene of interest
Foreign gene
Foreign cell
Bacteria have both Plasmid DNA and Bacterial DNA
Plasmids are used as cloning vectors in rDNA technology because they can
Replicate
transfer genes from one cell to another.
Bacterial transformation is when foreign DNA is transferred
Bacteria are able to take up new DNA very quickly, coming from other cells
Changing the Genome/DNA transfer = Bacterial Transformation
pGLO is a plasmid from a jellyfish that glows
They can put the plasmid in via Heat shock and Electric shock
Heatshock: By putting bacteria into very cold solution, plasmids are added, then in hot water the plasmid and bacteria combine, then in ice bath again.
pGLO, ampicillan etc. has an antibiotic resistance gene
Some of the bacteria will take up the plasmid but some will not
To find the recombinant plasmid, they use an antibiotic plate and the antibiotic resistant plasmid will form a colony and show that the plasmid has affected it.
A vector is a molecule used as a vehicle to carry genes of interest to foreign cell.
Bacterial plasmids are commonly used because of self replication etc.
These plasmids can be modified for specific usage.
PLASMID VECTORS
Multiple Cloning site - contains numerous recognition sites to allow gene insertion
Promoter - Initiates transcription
Origin of replication - for plasmid synthesis
Antibiotic resistance - selects modified cells
Reporter gene - makes products that attach to the protein to enable detection
STEPS IN PLASMID CLONING
Restriction enzymes to digest DNA sample and DNA plasmid
DNA ligase to seal
Transformation of recombinant plasmid into bacteria
Agar plates with selection antibiotic resistance allow bacteria with the resistance and gene of interest is cloned.
ANTIBIOTIC PLATE
The antibiotic plate is used to see if the transformation has occurred, as the antibiotic plate will kill any bacteria that has not been transformed, and transformed bacteria will grow because they have the antibiotic resistance and therefore show what has and has not been transformed.
Can be used to increase gene pool
Humilin
Insulin
Cloning Vector
A Chain - 21 Amino acids
B Chain - 30 amino acids
cDNA - copy DNA
‘S’ - suppression not resistant
‘R’ - resistant, immune
Fusion protein - A protein made from a fusion gene which is created by joining parts of two different genes.
Has 51 amino acids
Is a hormone produced by beta cells
Lowers blood glucose
Has a quaternary protein structure consisting of 2 peptide chains held together by disulphide bonds between cysteines
Chain A - 21 amino acids long
Chain B - 30 amino acids long
A plasmid vector and insulin gene are isolated from E.coli then it is cut open by the same restriction enzyme.
1 bacteria is used to make Chain A and a different bacteria is used to make chain B so that we have complete control over when chain A and B are together and manufactured.
Insulin will only become functional when its taken from the two cells.
When DNA is transcribe into mRNA then reverse transcription occurs its a DNA (single), polymerase is needed to make it double stranded.
Synthesis of Chain A/B
Obtain copy of the insulin A gene that is double stranded without introns
It must not have introns because bacteria do not have introns and cannot function with such.
Double stranded to anneal to the double stranded plasmid.
cDNA is copy DNA
pBR322 has ampicilin resistance and TET resistance. It will be used to insert the insulin A gene which will make the CHAIN A protein.
pBR322 is the cloning plasmid.
When only one restriction enzyme is used, the sticky ends may stick back together, so two restriction enzymes are used. EcoRI and BamHI
Decrease the risk/chance of the plasmid coming back together.
Now a recombinant plasmid that has ‘Insulin A gene’ and loses its TET resistance
TET is interupted
WHen rplasmid mix with E.coli, some of the rplasmid will be transferred to E.coli to make them transformed, but not all will be transformed.
Rplasmid - AMP(r) + InsulinA gene + Interrupted TET gene
3 POSSIBLE OUTCOMES
Bacteria with NO pBR322
Untransformed
NO resistance to tetracycline or ampicillin
Bacteria WITH pBR322
Transformed bacteria by plasmid
Resistance to Tetracycline and ampicillin
Bacteria with Recombinant pBR322 containing the insulin Gene:
Transformed bacteria by rplasmid
ONLY resistant to ampicillin.
Method 2 uses Beta - galactosidase gene that codes for B-galactosidase enzyme which breaks down lactose and glucose and galactose
It’s used as a marker to identify the plasmids that have the insulin A/B gene, by changing colour
B-gal is to next to the Insulin A gene and goes into.
Gene expression of this plasmid
Beta galactose
Insulin A gene
Resistance to ampicillin
3 POSSIBLE OUTCOMES:
Untransformed bacteria
Did not take any plasmid
Not resistant to ampicilin
Transformed bacteria
Resistant to ampicillin
Insulin Gene
No Beta Galactosidase gene → No fusion protein made
Transformed bacteria
Resistant to ampicillin
Insulin gene
Beta galactosidase gene - Fusion protein made.
To identify which colonies have functional insulin gene (ie: Beta-galactosidase gene) bacteria will be grown on agar plate with X gal.
X-Gal detects the presence of Beta galactosidase and insulin gene by causing bacteria; colonies to have a blue or white colour.
BLUE colony: Beta galactosidase & Insulin Gene present
WHITE colony: NO Beta Galactosidase and Insulin gene.
Ethics
Bioethics
Moral principles
Human rights
Welfare of people
Informed Consent
Ethics: A system of moral principles, right v wrong
Bioethics: Moral principles specific to biological science
Beneficence - must not hurt others, maximising benefits and minimise harm
People welfare is prioritised over interests of science
Informed Consent
Holding healthcare institutions accountable and reviewing scientist works
Unethical things;
Plagiarism
False reports
Dishonesty
Breaches in integrity
Strict guidelines for evaluation, publication and follow up
Ethical Approaches to Bioethics:
Consequence bases
Places central importance on the consideration of the consequences of an action (the ends)
Aims to achieve the maximisation of positive results, with the minimisation of negative results.
The focus is on the eventual outcome as opposed to the process taken to reach it
What is the end result? Does it outweigh the negatives of the process?
Does the ends justify the means?
Duty/rule base
Duty of Care
concerned with how people act (the means) and the process taken to get to the result.
places central importance on the idea that people have a duty to act in a particular way,
•and/or that certain ethical rules must be followed, regardless of the consequences that may be produced.
Ie: it is not acceptable to cause immediate or temporary harm in the pursuit of a potential ‘greater good’
Virtues based
Is person-based rather than action-based.
Consideration is given to the virtue or moral character of the person carrying out the action.
Providing guidance about the characteristics and behaviours a good person would seek to achieve to then be able to act in the right way.
Things that are self-seeking are unethical
Use keywords: Outcome, good virtues, process, means, etc.
Ethical concepts are used when identifying bioethical issues and are used to inform ethical guidelines
When deciding the extent to which the outcome of something or the course of action is ethically applicable
INTEGRITY:
The commitment to searching for knowledge and understanding.
Honest reporting of results, sources, in ways that permit scrutiny and contribute to public knowledge and understanding
Regardless of favourable or unfavourable results
Must be transparent
JUSTICE
The moral obligation to ensure fair consideration of competing claims
No unfair burden on a particular group from an action. Fair distribution and access to the benefits of an action.
BENEFICENCE
Maximising benefits
Minimising risks and harms in taking a particular course of action or position.
‘Zero harm’
NON MALEFICENCE
Avoiding causation of harm
The harm must be less than the benefits of the courses of action and outcomes.
Harm/Risk can be a little as long as it is outweighed
RESPECT:
Consideration of the intrinsic value of all living things
Regarding welfare, liberty, autonomy, beliefs, perceptions. Customs. Culture
Considerations for agency
When living things have diminished capacity to make their own decisions ensuring they are protected and empowered.
Photosynthetic efficiencies
Crop yields
CRISPR-Cas9
Bacteriophage
gRNA - guide rNA
sgRNA - single guide RNA
CRISPR is used for Gene editing to ‘find and replace’
CRISPR Cas-9 is an Endonuclease complex that naturally exists in bacteria to edit virus DNA (bacteriophage)
Clustered Regularly Interspaced Short Palindromic Repeats
CRISPR has two important features:
Palindromic repeats : short palindromic repeats
The spacers are segments of viral DNA (bacteriophages) that allow the bacteria to recognise the same virus in the event of subsequent invasions.
CRISPR can:
Repair the damaged DNA by using a template with the correct sequence.
Silence a faulty gene: The DNA can ‘self repair’ when cas9 cuts it; however, mutations can occur to introduce the ‘STOP’ codon.
Replace faulty gene sequence with the correct sequence, by using cas9 to introduce enzymes that would replace faulty bases with the correct bases.
Increase transcription of a gene by deactivating cas9 and adding activators.
Complement proteins do three things,
1. Stimulate phagocytes to increase phagocytosis
2. Highlight/Tag invaders for phagocytosis
3. Cytolysis the lysis of a pathogen forming Membrane Attack Complex (splitting of a cell)
Cytokines are signalling molecules
Peptide, protein or glycoproteins
A chemical signal for cells to carrry out immunologic responses
Released in response to cell damage or to indicate presence of pathogen
Crucial in controlling growth and activity of other immune system cells and blood cells
Trigger cells to;
Proliferate (reproduce)
Induce inflammation
Promote antibody response
Activate macrophage
Interferons (IF) are cytokines, protein and signalling molecules
Act as a warning signal from infected cells to all other cells
Viral host cell is triggered to synthesise interferon which targets neighbouring cells
Allergens are harmless substances NOT harmful substances
ANTIGENS & MHC
Antigen = Antibody Generator
An antigen is a unique molecule (eg. protein) that triggers the production of antibodies and the immune system
Could be part of a microbe or foreign substance (does not have to be whole pathogen); bacteria, pollen, protein
Immune response may be
Antigen non-specific → innate immune response
Antigen specific → adaptive immune response
The presence of antigens classify blood type on Red Blood Cells
Antibodies?
Antigens are foreign to organisms and stimulate antibodies that are specific to that antigen
One antibody will generally bind to one antigen
Antigens are non-self/foreign molecules that the immune system recognises as enemies.
Cells are self or non-self, non-self cells have antigens.
Self Markers (MHC) label the body’s cells as a friend and are tolerated by the immune system.
MHC = Major Histocompatibility complex
If MHC is present, immune cells will not be activated to respond.
MHCs are a group of venus that code for proteins found on the surface of cells that help the immune cells distinguish between self and non-self cells
Two main types in vertebrates: Class I and Class II
Class I markers are found on body cells, somatic cells NOT red blood cells.
Class II markers are found on immune cells, macrophages, dendritic cells, monocytes, B cells, antigen presenting cells.
Class I: All nucleated cells eg. Body cells
Act to identify cells self antigens from non-self antigens
Class II: Primarily on professional APC [Antigen presenting cells]
These cells present Antigen fragments on their class markers to T lymphocytes and other immune cells. Immune cells will have both classes EXCEPT RED BLOOD CELLS.
Professional antigen-presenting cells - APC, identify antigens as non-self and process antigens
Present the antigen on a MHC marker, so that immune cells lymphocytes so that they can identify and destroy/respond to invaders.
ANTIGEN PROCESSING:
Cell engulfs an antigen bearing particle
LYSOSOME FUSES WITH ENDOCYTIC VESICLE
Endocytic vesicle forms
Particle is digested nto bit
MHC markers bind fragments of particle
ANTIGEN MHC complex becomes displayed on cell surface.
Cytotoxic T Cells will target intracellular pathogens, cells that have been infected INSIDE the cell and if it is a body cell then the Cytotoxic T Cell will release cytotoxins and destroy the cell.
MHC I - Are from inside the cell
MHC II - Outside the cell/body, external.
ADAPTIVE IMMUNE RESPONSE
Third line of defence are the lymphocytes
Specialised Lymphocytes
B Cells
T Cells
Depending on where the stem cell lymphocytes are matured, (T cells in thymus and B cells in bone marrow) they will become T/B cells
Pre-T cells leave the bone marrow and go into the lymphatic system/ circulation and then go to the lymph nodes, which is where they wait for antigens
Two types of adaptive immunity: Antibody mediated immunity/Humoural Immunity and Cell mediated Immunity
B cells → Humoral Immunity/Antibody Mediated Immunity → Plasma Cell which makes Antibodies, B CELLS do not make antibodies only the plasma cells.
T Cells → Cell Mediated Immunity → matures in the Thymus glance
Cytotoxic cells/CD8+ - will release cytotoxins
Helper T cells/CD4+ - will release cytokines
T Memory - A memory cell that will become either cytotoxic or helper T cells, for future infections
Regulatory Cell - T Supressor, stops the fighting
Natural Killer Cells - Lymphocytes - Not part of the adaptive immunity response, part of the innate immunity
B Memory - A memory cell that will become a plasma cell again
Cytotoxic cells have the CD8 glycoprotein and are activated by APC’s (antigen presenting cells)
They defend against intracellular bacteria, viruses, cancerous cells, transplanted foreign tissue, protozoa, fungi and worms.
Activated by MHC Class 1, releases cytotoxins such as perforin, granzymes to induce apoptosis.
Helper T Cells are not cytotoxic or phagocytic
They activate humoral immunity: B cells to make plasma cells to make specific antibodies
Cell mediated immunity: TC cells to release cytokines
Innate immunity: macrophages to initiate phagocytosis.
A naive Th-Cell means it hasn’t come in contact with that specific antigen before
Plasma b cells contain LOTS of rough er and undergo apoptosis
They make antibodies and antibodies are proteins so they need rough ER because ribosomes make proteins
Antibody Structure: Globular glycoproteins, quaternary structure
They are immunoglobulin
Two heavy and two light polypeptide chains (4 in total)
Chains are held together by disulphide bridges with Variable and Constant regions.
Order of amino acids determines the shape of the variable region binding site
Antibodies can neutralise antigens, agglutination (clumping bateria up), precipitation, assist in complement protein, enhance phagocytosis, inflammation, cell lysis
5 immunoglobulin isotypes
Humoural immunity
B cell receptor is the antibody
A naive B cell that has just processed the same antigen will present the antigen on its receptor
Lymphocytes
Circulatory system
Lymph nodes
Lymphatic vessels
Lymph fluid
Capillaries
The lymphatic system protects from infection and disease
Part of the immune system
Lymph fluid pass through lymph nodes
Network of lymph vessels connects the lymph nodes together
Lymphatic system acts as a one way drainage system, transports fluid from body tissue, houses lymphocytes and filters cellular waste
Capillaries are necessary so that all cells have access to oxygen and to get rid of CO2
Lymph Nodes, small structures that work as filters for harmful substances, located in strategic points
Immunity
Symptoms
Innate Immunity
Humoral Immunity
Cellular Immunity
Immunisation - harmless part of microbes are introduced to trigger the body’s immune response
Herd immunity
Vaccine - A suspension of antigens that are deliberately introduced into the body
Immunity can be
Innate or Adaptive
Natural or Artificial
Active or Passive
Natural Immunity occurs through contact with a disease causing agent, it is not deliberate; by chance
Artificially acquired immunity develops only through deliberate actions such as immunisation.
Both natural and artificial immunisation have the same result of activating adaptive immune response
Passive immunity is acquired through the transfer of antibodies, their immune system has not been activated, they are NOT receiving antigens, but instead antibodies. Memory cells are not made, they only have antibodies. Immunity only lasts as long as the antibodies are present.
Active immunity is the adaptive immune system activated. Delivery of antigens.
ACTIVE-NATURAL IMMUNITY
Pathogen/Antigen enters body (illness)
Adaptive immune system is activated
Memory cells are made
Long Term Immunity
ACTIVE-ARTIFICIAL IMMUNITY
About immunisation (vaccines)
Herd immunity
Active, deliberate process
Antigens are introduced and memory cells are made
Long term immunity
Types of Vaccines: Subunit or Whole agent vaccines
Subunit - contains some part or product
Whole-agent - contains whole, non virulent microorganism which can be inactivated (killed) or attenuated (weakened)
Herd Immunity, the resistance to the spread of a contagious disease within a population that result if a sufficiently high proportion of individuals are immune to the disease especially through vaccination
Minimisation of an epidemic
PASSIVE ARTIFICIAL
Antibodies injected
Used when a very rapid immune response is needed like antivenom
Human antibodies are injected
ANtibodies come from blood donors who recently had vaccination
Only provides short term protection
No memory cells
PASSIVE NATURAL
Mother’s antibodies pass across the placenta to the foetus
Colostrum (the first breast milk) contains lots of IgA which remain on the surface of the baby’s gut wall and pass into blood.
Immunotherapy
Metastases
Benign
Malignant
Cancer
Cancer is a group of diseases involving abnormal cell growth, with the ability to spread throughout the body
Benign or Malignant tumours
Beninghn cannot spread by invasion or metastasis
Malignant can spread through the body via the bloodstream
Immunotherapy uses Vaccines and monoclonal antibodies (mAbs)
Tumour cells thrive because they are able to hide from the immune system by expressive defective class-1 MHC
Immunotherapy uses the host system
SPECIFIC:
Marks cancer cells so immune system can find and destroy
Triggers response
B&T lymphocytes are stimulated to target cancer cells
Contain: Peptides, antigens or whole proteins of cancer cells and adjuvants
Adjuvants: substances that enhance the effect of a vaccine or other treatments
No side effects
Classified: Preventive or Therapeutic and personalised
Preventative, harmless virus-like particles containing viral DNA trigger immune response to create antibodies and memory cells etc.
Therapeutic: For the treatment of someone experiencing cancer. The tumour is not yet recognised by the immune system so immune cells are trained to recognise antigens and injected into individuals so the adaptive response is activated to be able to recognise and attack the tumour.
Monoclonal Antibodies are made to treat diseases
mAbs target specific antigens found on diseased or cancerous cells
mAbs can act directly when binding to cancer specific antigens to ;
Induce immunological response: apoptosis
Highlight cancer cells to immune cells
Block growth signals
Deliver toxins
NON SPECIFIC:
Boot immune system to work better
Trigger innate response, cytokines et: interleukin and interferon
AUTOIMMUNE DISEASES:
Diseases that have the immune cells targeting self-cells
Cytotoxic T cells, B cells activating, mast cells→ histamines and inflammation
No cure
Immune suppression that can help control overactive immune response and decreasing pain
Anti-inflammatory
Alongside immune suppressive medication
Acts as competitive inhibitor signals.
Interleukin competitive inhibitor, stops signals for immune system to work
Can stop the b cells, and t cells etc. Targets the immune system that will be targeting self-cells
Bacterial resistance
Antigenic shift
Pathogens
Population
Evolution
Reservoir
Novel Strain
Rational Drug Design - Targeted approach to designing new drugs, involves analysing the structure of a pathogen
Gram-positive - thick cell wall of peptidoglycan
Gram-negative - thin cell wall of peptidoglycan
Broad spectrum antibiotics - targets wide range of bacterial species
Narrow spectrum antibiotics - targets one or two bacterial species
Bacteriostatic
Bactericidal
Microevolution
Antigenic Drift
Antibiotics
Bottleneck Effect
Predation
Evolution = Change
Outbreak: the occurrence of one or several cases of a disease in an area in which it is not normally present
Epidemic: an uncontrolled outbreak that is the infection of many people simultaneously
Pandemic: An epidemic on a global scale, disease spread worldwide
Endemic: A disease that exists permanently in a particular region/population
Outbreaks require identification of cause, treatment, prevention of spreading and another outbreak
Condition for a pandemic
New pathogen/Novel strain - there is a lack of interaction with antigens and cannot be protected against because there is no immunity, no vaccine
Pathogen infects people and non-human hosts providing a ‘reservoir’
Pathogen is easily transmitted through direct contact, air or vector
Infected individuals are not isolated
No vaccination or preventative measures are in place
No control measures (masks, quarantine)
Influenza virus Structure
Two spike proteins, neuraminidase/sialidase and hemagglutinin
Nucleoprotein - RNA
Neuraminidase - enzyme, viral exit - cuts out
Hemagglutinin - viral entry, receptor - goes in
The virus enters and undergoes endocytosis, the nucleus is usesd for mRNA synthesis and RNA replication so now the virus has taken over the cell and will be used to make the virus while the virus exits and infects more cells
Subtypes of Influenza A are differentiated on the basis of the two surface antigens
Three Subtypes of H (Hemagglutinin) (H1,H2,H3)
Two of N (N1 and N2) generally cause the annual epidemics
Influenza A viruses are classified by A,B,C
Influenza A can cross species
A&B are main causes of epidemics/pandemics Type C cause mild versions
ANTIGENIC DRIFITING
There can be gradual minor changes in HA/NA caused by point mutation
Occurs in A&B
Vaccines are annually updated
ANTIGENIC SHIFT
Sudden Major Change, genetic reassortment of genes caused, when two subtypes infect a host, direct transmission from other animal to human introduces a NEW novel strain
Explosive Spread
Only in Influenza A
Creates a pandemic/epidemic
What are the scientific and social challenges presented in terms of the treatment strategies and vaccine programs (caused by viral antigenic drift and shift)?
Scientific: Vaccines created in response to viruses would have to change dramatically to accommodate for major changes due to antigenic shifts in the virus’s structure. Identification of mutation would be difficult and the difficult within identifying antigenic shift or drift.
Social: Immune responses of the population are naive and there is no immunity to a novel strain as the antigenic shift has drastically changed the virus’s structure. Vaccination development can be expensive and time-consuming as well as how measures will be implemented such as quarantines, lockdowns, masks, vaccinations etc.
CONTROL:
Antiviral drugs that prevent viral entry by binding to receptors
Inhibition of enzymes that catalyse reproduction of virus genome
Blocking transcription and translation
Prevents viruses form leaving cells to prevent further infection
RATIONAL DRUG DESIGN:
Rational Drug Design - Targeted approach to designing new drugs, involves analysing the structure of a pathogen
Uses this information to design a drug that will mimic or block the action of the disease-causing agent.
Produces drugs that have complementary shapes to the active sites of the pathogen or molecule they are targeting.
OTHER CHEMICALS TO CONTROL PATHOGENS:
Disinfectants - Non specific,
Antiseptics - Non specific, against bacteria, viruses and fungi
Antibiotics
Antibiotics:
Substances produced by microorganism or artificially that in low concentrations inhibits the growth or kills microorganisms
Broad spectrum antibiotics - targets wide range of bacterial species
Negative: Unnecessary introduction of antibiotics that may cause harm
Narrow spectrum antibiotics - targets one or two bacterial species
Better but takes time
Bacteriostatic - slows growth of bacteria by interfering with synthesis processes, like DNA replication, enzyme activity or protein synthesis
Bactericidal - kills bacteria, as an example may prevent the growth of cell walls to kill the bacteria.
Antibiotics only for bacteria because viruses hide in body cells and also they don’t have cell walls so bacteriostatic can’t target them, bactericidal can’t destroy the cell wall; they have different structures to bacteria
Four types of antibiotic resistance
Impermeable barrier, target modification, antibiotic modification, efflux pump mechanism
Evolution = Change in allele frequencies
Bacteria transfer genes easily and go into plasmids/DNA, high reproduction rate, exponential growth of population
Population: A population is the number of all the organisms of the same group or species, which live in a particular geographical area, and have the capability of interbreeding to produce fertile offspring.
Species: A group of living organisms consisting of similar individuals capable of exchanging genes or interbreeding.
Alleles: Alternate forms of a gene, two alleles is 1 genotype
Allele pool/Gene pool: The sum total of allele for all genes present ina population at one time
Large gene pool indicates genetic diversity and biological fitness
Small gene pool indicates low genetic diversity and biological fitness → Increasing chances of extinction
Used to determine allele frequency proportion of a particular allele within a population
Biological Fitness: The level of fitness a species has for surviving changes in environment based on the ability to adapt to new situations
Evolution change in allele frequencies over time
Mutation
Random change in the genetic composition due to changes in the DNA base sequence or chromosome
Point mutation
Gene Flow
Movement of alleles into or out of populations due to immigration/emigration
Gene flow keeps separate populations similar
Sexual Reproduction
Sex can introduce new gene combinations
Alter allele frequencies if mating is assortative
Random mating
Non-random mating
Assortative Mating - Preference for similar genotypes/phenotypes
Disassortative Mating- Preference for different genotypes/phenotypes
Genetic Drift
Completely random/chance
Causes the allele frequencies to drift from one generation to the next
No selective agents
May cause gene variants to completely disappear
Genetic drift has a greater effect on small populations (5/100 vs 5/10)
Population bottlenecks and Founder Effects
Bottleneck → Catastrophic event reduces population and reduces genetic diversity by chance
Founder Effect → Small group moves and reproduces in a new location, genetic variation is low
Gene Flow | Genetic Drift | |
Occurrence | Occur thru migration from one to another pop | Occur through random events. |
Population size | Larger population | Smaller population |
Reason | Inbreeding or inbreeding through migration | Sudden change or sampling error |
Evolution | Through migration | Thru bottleneck/founder |
Natural Selection/Selection Pressure
Change in gene pool composition as a result of differentially selective environmental pressures
Predation, Abiotic factors, Nutrition, Disasters, Finding a mate
Selective pressures are biotic and abiotic factors that select for certain characteristics in a population to be passed on and selected against other characteristics that will not be passed on
Eg. Dark bug camouflaging on a dark tree doesn’t get eaten by bird, bright coloured bug gets eaten
1. Variation: Intraspecies differences
2. Selection Pressure: A struggle is applied to population
3. Adaptations: Survival of the fittest
4. Reproduction: Adaptive quality is passed onto offspring
5. Change in population: Allele frequency/microevolution
Increase offspring of a certain trait due to survival of the fittest (organisms most capable of reproducing produce more offspring with whatever trait they have that has enabled them to survive and reproduce)
If there is no selective pressure, different traits do not matter
There needs to be a struggle to live (selective pressure)
Selective pressures:
Just because a phenotype has been wiped out does not mean the allele has disappeared/genotype may still have the alleles that will trigger the recessive allele
The allele cannot be dominant because then even a single allele will trigger the phenotype thus, if the allele IS dominant it has gone extinct
NO SELECTIVE PRESSURE NO CHANGE!
Things that mean NO change to allele frequencies: Large population, no mutation, no migration, random mating (organisms choose partners randomly), no selection (no traits can be favoured)
Point mutation
Block mutation
Chromosome
Meiosis
Allele
Gene sequence
Mutation
Substitution
Deletion
Addition
Non-disjunction
Adds a new allele to increase gene pool
A mutation is a change in the gene sequence or chromosome.
Gene mutations (point mutation) localised changes to DNA base sequence ie: substitution, deletion or addition
Chromosomal Mutations (Block mutation) large scale mutations occurring during meiosis that can change chromosome structure and number
Gene mutation:
Missense substitution
Type of mutation is a change in one DNA base pair resulting in amino acid subbing in for another in the protein made by a gene
Eg. Sickle Cell Anaemia
Nonsense Substitution
Accidental mutation codes for STOP codon resulting in a shortened protein/junk protein
Insertion (Frame Shift)
An insertion changes number of DNA bases, protein may not function properly as point mutation has caused a frame shift
Silent mutation:
Nothing is changed, redundancy allows for the same amino acid to be produced
Deletion (Frame Shift)
Changes number of DNA bases, frame shift because protein will be changed sequence
Frameshift Mutation
Can change every amino acid that follows point of mutation and alters a protein so much
Changes the sequence, shifts reading frame
Chromosomal Mutations [Block Mutation]
Block mutations can cause polyploidy, changes in chromosome number and structure.
Duplication
Alleles are duplicated/added
Inversion
Positions are swapped, frequency doesn’t change but sequence does
Deletion
Removed segments of chromosome
Insertion
New alleles introduced
Translocation
Also swaps but with a different chromosome
Meiosis:
Meiosis can stuff up and nondisjunction may occur, Main cause for aneuploidy
Aneuploidy, incorrect chromosome number
(Chromosomes do not separate properly)
Variation is introduced because random alignment and crossing over/random assortment when there is exchanging of whatever
Polyploidy cells are organisms containing three or more times the haploid number of chromosomes like 3N or 4N, uncommon in animals, common in plants
Occurs through allopolyploidy and autopolyploidy
Allopolyploidy - An individual or strain whose chromosomes are composed from two different species to produce hybrids
Can have the full chromosome set of two different species
Autopolyploidy - resulting in offspring with two sets of chromosomes from it’s own species
Criteria: variation in a population and heritable traits
Humans select
Determine desired trait
Interbreed parents with desired trait
Select offspring with desired trait and interbreed them
Process continues until reliable reproduction of desired trait is achieved
Problems with selective breeding
Gene pool has been reduced and alleles are lost therefore reduced resistance to environmental change
Reduced Biodiversity Genetic diversity decreases, ability for adaptation is decreased
Increased genetic abnormalities, Genetic defects can be selected for with favourable traits
Stratigraphy
Geologic Time Scale
Palaeontology - the study of fossils
Body fossil
Trace fossil
Impression fossil
Mineralised fossil
Intermediate
Transitional fossil
Permineralisation
Index fossil
Sedimentary
Strata
Geologic Time Scale (GTS) is a system of chronological dating that relates geo strata (stratigraphy) to time
First prokaryotic life form - Cyanobacteria
Fossils preserved in rock, soil or amber
Remains of organisms
Palaeontology is the study of fossils
Lowest rock layers are older
Rock layers formed later contain more complex organisms
Variety also increases
Body Fossils - fossilised remains of an organism eg: bones, leaves
Trace Fossils - No parts of an organisms, impressions of activity eg: footprints
Impression Fossils - Organism decays and leaves an impression the rock/earth
Mineralised fossil - minerals replace the organism structure
Fossilization requires
Rapid burial: protection against scavengers, erosion and damage
Low oxygen: protection of oxygen damage and lack of decomposition
High pressure: to promote mineralisation of remains
Remains undisturbed: To allow for permineralization
Hard body parts: eg. teeth, shell
Death and decay: Soft body parts decay, leaving only hard body remains
Deposition/rapid burial - hard remains are rapidly covered with silt and sand and layers build over time
Permineralization - pressure from layers od dirt and rock cause hard organic material to be replaced by minerals
Erosion/ exposure - movement of earth plates may displace the fossil and return to discovery
Soft body fossils are less likely because they have more water and are more likely to decompose, more likely to create impression fossils (ie: jellyfish)
Transitional fossils: Fossil that can link two different lifeforms, remains of a pre-existing organism that shows a progression/transitions
Should show transitional/intermediate characteristics
REMINDER: GO THROUGH PRESENTATION 6. FOSSILS WE LOST TIME AND GO THRU IT BETTER
Absolute dating: anything that gives exact dates and numbers, numerical dating, determined by radiometric dating, estimates the age in years by measuring certain radioactive isotopes the object contains.
Relative dating: estimates the age of fossils found within strata, cannot tell the actual age of the fossil, using index fossil/stratigraphy, older than this/that, MUST COMPARE SOMETHING
Sedimentary rocks
Some of these layers may be laid down by water (Sedimentary) or volcanic activity (igneous)
Importance of the sequence in which is was deposited etc.
Relative dating is important to figure out index fossils
Index fossils are organisms that were geographically widespread and abundant but only existed for a limited span of time.
Must be distinctive, globally widespread and recognizable, became extinct quickly to pinpoint precise time periods
ABSOLUTE DATING
Radiometric:
Using the isotopes of carbon to determine age
Isotopes: Variation in neutrons
The presence of Carbon-13 and Carbon-14 indicate how long something has been around and the decay of that isotope
Isotopes go through radioactive decay
Parent Isotope is unstable
Daughter Isotope is stable
Radioactive decay is when the parent isotope becomes stable
When half of parent isotopes have decayed/become stable becomes 1 half life and so forth
Carbon-14 is not old enough for REALLY OLD shit
Carbon 14 takes 5730 years for half of the isotopes to become stable and become N14
Process of C-14 → N-14
Cosmic radiation heat N-14
N-14 loses a proton → C-14
C-14 + C-12 are in the atmosphere and get absorbed by living organisms
When those organisms die, bones lose C-14 as it becomes N-14 via beta decay (gains a proton)
Maximum limit of this method is 60 000 years
LIMITATIONS OF FOSSIL RECORDS
Organisms decompose rapidly
Are eaten
Soft-bodies organisms do not fossilise easily due to water
Small fraction of organisms die in conditions favourable to fossilisation
Fossils are still unearthed
Macroevolution
Speciation - the evolution by which new biological species arise over time
Species - Organisms that can produce fertile offspring with one another excluding asexual reproducing organisms
Ancestral population
Allopatric Speciation
Prezygotic Barrier
Postzygotic barrier
Geographical isolating mechanism
Isolating mechanism
Reproductive mechanism
Adaptive radiation - divergence of a large number of related species from a common ancestor
Speciation is MACROevolution - the evolutionary process by which new biological species arise over time
Ancestral populations are divided then isolated preventing gene flow
Different selective pressures will create differences in population
Allopatric speciation
Ancestral population: There is gene flow and variations
Isolating Mechanism: prevents gene flow (the movement of genes between populations)
Geographical isolation
Reproductive/Genetic isolating
Mutation creates new variants in different areas
Natural Selection: different selection [pressures select for new vairants
Speciation: individuals from each population can no longer produce fertile offspring with each other or the original ancestral population.
Reproductive Isolation
Mechanisms that prevent mating and reproduction, ie: individuals not responding to courtship
No gene flow
No exchange of alleles between populations
Prezygotic isolation
No fertilisation → No zygote
Postzygotic isolation
Zygote is formed but it is inviable or infertile (dies or can not reproduce)
Pre-Zygotic
Post Zygotic:
Adaptive radiation - divergence of a large number of related species from a common ancestor
CHARLES DARWIN'S GALAPAGOS ISLAND FINCHES:
Finches with different features found on Galapagos islands all descendants of the mainland ancestral species.
Geological Isolation
Sympatric Speciation
Species share the SAME geographical area but are reproductively isolated
Isolation comes from within the group
Assortative mating
Relatedness
Relation
Divergence - Speciation
Homologous Structures
Adaptive radiation
Vestigiality
Molecular homology - sameness on a molecular level
Comparing anatomical structure, Mitochondrial DNA, genetic sequences
Structural vs Molecular
Homologous Structures
Evolved from the same structure in an ancestral species
Different functions
Immediate common ancestors
Organisms show divergent evolution
Adaptive radiation
Eg. Pentadactyl limb
Vestigial - Describes homologous characters of organisms which have seemingly lost all or most of their original function in a species
Vestigiality can be structures, behaviours and biochemical pathways
Changes to the environment have rendered these structures redundant and so over time they have lost their functionality
Same function
Different structure
No immediate common ancestor
Organisms develop the same structure due to convergent evolution
Molecular clock hypothesis is:
Changes in DNA and proteins are constant over evolutionary time and across different lineages
The amount of molecular change between two species measures how long ago they shared a common ancestor
Molecular clock calculations are carried out on DNA or amino acid sequences btw species to establish relatedness
Comparison of DNA sequences and amino acid sequences
If divergence has occurred further back in time, there will be less similarity/less molecular homology
Mutation Rate:
Change in DNA over time
Can be expressed as the number of nucleotide changes over a million years
Molecular clock uses the rate of accumulation of mutations of DNA to determine how long ago divergence occurred.
Less Related Species:
More mutations
Greater differences
Divergence occurred further back in time
Closely related species
Less mutations
Less differences
Divergence occurred more ‘recently’
Amino acid:
Differences in amino acid sequence reflect changes in DNA sequence
Changes in the gene nucleotide should build over time
DNA comparison:
Direct Comparison of DNA base sequences
Comparing whole genome
DNA hybridisation
Comparing karyotype
Mitochondrial DNA
Mitochondrial DNA
Only inherited from mother
Mostly for recent (20 mil)
Can be recovered from teeth and bones
There’s a lack of recombination of mtDNA, remains the same
Higher mutation rate: Contains non-coding regions known as the D-loop that mutates at a higher rate
High copy number - cells have lots of mitochondria
Maternal inheritance: mtDNA is inherited from the mother only, to establish ancestry • It is used as a molecular clock.
Mitochondrial Molecular Clock: rate at which mutations have been accumulating in the mitochondrial genome of hominids during the course of human evolution.
Closely related species will show more similarities in base sequences, genome, DNA, Chromosomes, mtDNA
Lineage
Phylogenetic
Ancestor
Ancestral lineage
Descendant
Relatedness - determined by comparison of aa sequences or DNA to establish phylogeny, how recently species diverged
Phylogeny - refers to an evolutionary line of descent, can be determined by comparing sequences in different species.
Phylogenetic trees act as evidence for relatedness
Phylogeny refers to the evolutionary line of descent, determined by comparison
Phylogenetic tree:
Hypothesis relatedness
Phylogram
Compares sequences that have a constant rate of mutation (evolutionary clocks)
Mitochondrial DNA is a useful source as it is maternally derived has a known mutation rate and lacks recombination
Difference → mutation in either nucleotide of amino acid sequences.
Evolutionary trees can evolve to alter hypotheses if new evidence alters understanding
All living organisms have Cytochrome B
Hominidae
Genus
Family
Order
Species
Hominoid
Primate
Primary ancestors would have had
Arboreal
Grasping hands
Long, mobile limbs
Quadrupedal locomotion
Binocular vision
Upright sitting position
Nails instead of claws
Large eyes to improve eyesight, colour vision
Large highly developed area associated for vision
Reduced development for smell
Different types of teeth for wider variety of food sources
Singular birth: Longer parental care, increased infant dependency
Tails
Hominoidea - superfamily that includes apes and humans
Hominoids - members of the superfamily hominoidae
Hominids - all modern and extinct great apes. Gorillas chimps etc. and Immedieate ancestors
Hominins - any species of early human that is more closely related to humans that chimpanzees including modern humans
Pre-Hominins - Arboreal lifestyle, food resources were readily available in near-continuous forest
Cooling climate —> Trees became scarce
As trees became scarce, pre-hominins were forced to leave trees in order to seek out food sources
Bipedal Walking
S-curved spine
Inward femur angle
Pelvis shape
Foot shape/structure
Reduced Canines (Not related to walking)
Foramen Magnum - hole in skull
Brain size/Skull
Bipedal motion through S-shaped, flexible spine for balance
Femur/tibia angled inwardly for centre of gravity and allow for balance and walking
Short broad pelvis to allow for attachment of large powerful muscle
Bow shaped to support torso organs
Chimpanzee feet has opposable thumbs
Human feet has an arch, acts a spring, loss of big thumb, slightly larger toe
Transverse Arch - Converts foot into a spring allowing for transmission of stresses and improving walking efficiency ef
Foramen Magnum is the hole at the base of the skull thru which the spinal cord passes
If the foramen magnum is positioned towards the back (posterior) - quadruped
Foramen is more centred/to the front – Bipedal
Larger brain size, reduced brow ridge and flatter face
Cranium Capacity - Mass of brain that can fit into a human
Trends in Skull Anatomy
Shape and slope of forehead
Brow ridge
Facial Angle
Size of teeth
Protrusion of mouth
Position of foramen magnum
Size and shape of zygomatic arch
Brain case - size and shape
Size of mandible
Sagittal crest present? (Humans, not present)
Shape of occipital region
Differences between skulls of Australopithecus Afarensis and Homo Sapiens
Differences | Austra | Homo Sapiens |
Size of Skull | Smaller skull
| Larger skull |
Brow Ridge | Has one | Doesn’t have one, not very prominent |
Foramen Magnum | In the posterior, towards the back to allow for quadrupedal posture | In the anterior, centre of skull so head is atop spine and balanced |
Teeth Size | Larger teeth
| Smaller teeth |
Jawbone | Wider Jawbone | Narrower Jawbone |
Face Slope | Slanted slope, on an angle, teeth are jutted out and jaw recedes at an angle | Protruding jaw but straight face slope/vertical |
Zygomatic Arches | Very prominent cheek bones | Reduced |
Arms | Longer than legs, used for walking | Shorter than legs, not used for walking |
Why was Bipedalism Selected for/Adapatations!
Enabling a more proficient use of tools by freeing hands when upright
Bipedalism is more energy efficient
Thermoregulation: Lowers body temperature as solar radiation is retatined
Greater view of surroundings therefore able to escape predators more effectievly
Greater ability to disperse and cover more ground leading to habitat variability
More effective mating strategies leading to successful reproduction
Reduced canines was also selected for
Ability to carry food and weapons when walking
Carrying offspring while moving and eating
Hair loss
Retention of head hair for reflection of heat
Easier to control parasites
Thermoregulation - Less trapped heat, Greater heat loss, well developed sweat glands
Early Hominins
Australopithecus
Homo genus
Environment, what they looked like, how they moved etc.
Homo Erectus
First human emigrant
Left Africa
Founder effect, some remained in Africa
Larger brains and advanced toolmaking
Left btw 100 000 and 1.6 million years ago
Wore skins enabling travel from Africa into china and south East Asia
Homo Floresiensis
‘Hobbit’
Small stature
Wide pelvis and hunched shoulders
Lived in Asia between 100,000-60,000
Flat face’
Theories
Smaller body possibly due to surviving with constrained resources, island dwarfism
Descendants of homo habilisi
Has a pathological condition in modern humans (microcephaly)
Homo Neanderthalensis
Complex species
Bigger brain than humans
Similar but they have a bigger brow ridge
Shorter
Had a more robust skeleton and muscular
Cousins NOT direct ancestors, shared a common ancestor
Co-existed with modern humans
Went extinct 28000 years ago
Shows many single base differences however DNA is very very similar
Neanderthals did not contribute any mitochondrial DNA to any homosapiens living today
Human females and Neanderthal males
0% of Neanderthal DNA in African populations
1-4% in people of European or Asian descent
Did not migrate to Africa, only came in contact in Eurasia
Possibly went extinct due to transmission of disease, pressure of incoming migrant humans, or being slaughtered by modern humans (killer ape theory)
Homo Denisovans
Diverged from modern humans about 500 000 y.a., related to neanderthals
Diverged from neanderthals about 300 000 years ago
Possibly resembled neanderthals
Epigenetics:
adding methyl groups, not actually altering DNA etc
Working out pattern of methl or whatever could see gene expression and effects on appearance.
Archaic Human
Includes neanderthals, H. Floresiensis and H. Denisova
Mainly differed in skull
Backward sloping forehead
Big brow ridge
Long elongated skull
Why did they leave Africa:
Depletion of resources
Competition for resources
Climate change - droughts leading to starvation
Curiosity
Human Migration Evidence
DNA Evidence to support the hypothesis that early hominins migrated out of Africa around 150 000 ya
DNA evidence suggests interbreeding btw modern homninins and nanderthals in Europe and the Middle East
Reached australia 35-60 000 ya
The Pleistocene Ice Age created a land bridge that connected Asia and Alaska over 13,000 years ago.
DNA suggests that modern humans reached australia continent approx 55 000 ya
Out of Africa = Replacement theory
H. Erectus left, evolved into Archaic humans
H. Sapiens left 200 00 years later and replaced archaic humans
Out of Africa model II = Assimilation theory
Same as A except some H. erectus survived and interbred and joined H. Sapiens
Multiregional - Continuity theory
• Significant migration of H.erectus across Africa, Asia and Europe for the last 1.8my. • Isolation of the populations à divergence of gene pools, traits & behaviour. • interbreeding btw populations could have occurred • genetic drift led to the DNA of the other species being lost from the H.sapien genome.
Multiregional theory 2
Haplogroup
Group who share common ancestor on paternal or maternal line
Inherited Y chromosome
Inherited mtDNA
Indigenous Migration
Single migration from Africa to Australia 60 000
42 000 ya, extinction of megafauna
Aboriginal people have strong sense of country and place
Place- space mapped by intangible boundaries that individuals or groups of Torrest Strait Islander peoples occupy and regard as their own via spiritual and emotional connections
Connections:
Connection via Maternal and paternal lines of descent • Clan group • Language groups • Spiritual connection
VCE Biology 3/4 Notes
Procedure - The set of processes used to define the scientific method typically including a hypothesis, experiment, observation, analysis, conclusion and evaluation
Hypothesis - If the (DV) then the (IV)
Independent variable - Variable that is manipulated and has a direct effect on the DV
Dependent variable - the measured variable that is observed/recorded
Repeatability - The ability of the experiment to be repeated identically in the same environment
Precision - The degree to which the results are consistent
Accuracy - The degree to which the results are close to the true value
Reproducibility - The ability of the experiment to be repeated with the same materials and method in a different environment
Validity - The experiment is measuring what it's supposed to be measuring
Bar Graphs - Different groups of data
Line graphs - Ranged set or continuous data
Control set up - The experiment without the independent variable
Controlled variables - Variables that are used to keep the experiment and control set up the same except for the independent variable.
Procedure - The set of processes used to define the scientific method typically including a hypothesis, experiment, observation, analysis, conclusion and evaluation
Hypothesis - If the (DV) then the (IV)
Experiment
Observation
Analysis
Conclusion
Method
Investigation
Independent variable - what is manipulated and has a direct effect on the DV
dependent variable - the measured variable that is observed/recorded
Experimental Set-up - To test the hypothesis, will have the independent variable
Control Set-up - Used to compare and confirm findings in experimental set-up, Does not have the IV,
Controlled Variable - Factors that are applied in both setups. Constant factor. purpose is to ensure that only one variable is tested at a time in an investigation: To make it valid. Eg. how much water you drink, weather, light, etc.
Repeatability
Precision
Accuracy
Reproducibility
Set of procedures to gain knowledge
6 key steps
Question
Hypothesis: educated guess or prediction, must be testable, states variables, “if, then”
Experiment
Observation
Analysis
Conclusion: statement of whether the original hypothesis was supported or refuted
Steps are not always linear but systematic
To make a hypothesis: ask a research question then change that into hypothesis
VCAA formula: If (DV) (relationship to IV) then (trend/effect on DV) and (IV whatever)
Controlled variable ≠ control set up
An investigation should only have ONE independent variable
Two setups only: Experimental Set-Up + Control Set-up
Results will be in graphs or tables.
RULES:
Title your table, including IV and DV
Column headings - Units included in Headings
Left column must be the IV
Data entry must have the same number of decimal places
Keep things neat and aligned
TAILS
Title
Axis
Interval
Label
Scale
Bar graphs are for different groups of data
Line graphs are a ranged set of data/ continuous data
Don’t forget repeatability
Example
Observation: Mice that are given Vitamin D increase the Calcium Absorption
Independent Variable: Vitamin D being given to mice
Dependent Variable: The Calcium absorption in the mice blood
Hypothesis: If Mice are given Vitamin D then the Calcium absorption in their blood will be increased.
OR
If mice are given Vitamin D then the Calcium levels in the blood will be less than the mice not given Vitamin D
Experimental set-up
100 Mice given Vitamin D
Vitamin D
Controlled Set up
100 Mice not given vitamin D but instead a placebo
Controlled Variables:
Amount of Vitamin D and placebo
Duration
Species of mice
Age of Mice
Same period of testing
Same food
Same water
Mice Sex
Results to support hypothesis:
A greater % of mice in the experimental group will have less calcium the blood while the control group show no increase of calcium absorption
Results to refute hypothesis:
A greater % of mice in the experimental group show no change in calcium levels
Accuracy
Not a quantity
How close to the true value
Bottle weighing 3kg measuring at 2kg is inaccurate
More trials does not increase accuracy
Precision
How closely two of more measurements agree with each other
Little spread among the values
If true value is 2.4 and results are 2.3, 2.3, 2.4, 2.4 then accurate and precise
5.6, 5.7, 5.9, 5.8 precise not accurate
Lack of accuracy but precision is systematic errors
Lack of precision could mean random errors
More trials can increase precision
Repeatability
Same method on identical materials with the same condition
The exact same experiment with the same person who did the experiment or operator
To obtain same results
Provides evidence of validity and reliability
Reproducibility:
Same method, same materials but different conditions
The exact same experiment in different environment with possibly a different operator
Identifying random or systematic errors
To obtain the same results
Validity
A measurement is valid if what is being measured is exactly what was claimed
Data is valid if the independent variable is the ONLY variable
Similar to accuracy
Reliability
Similar to precision
Random Errors
Random errors are unpredictable events or mistakes that affect the results
Outliers
Mistakes etc.
Affects accuracy and precision
Systematic Errors
Occurring the measuring system and affects every single measurement
Does not necessarily affect precision, affects accuracy
Re-calibrate machines
U3AOS2
Enzymes → P1
Photosynthesis → P2
Respiration → P3
Catalyst
Proteins
Enzymes
Enzyme functions
Catalyse
Substrates
Coenzymes
Active Site
Enzyme substrate complex
Enzyme product complex
Anabolic - building up, 1 + 1 → 2, always requiring energy, (endergonic)
Catabolic - breaking down, 2 → 1 + 1, reduce complexity, always releasing energy (exergonic)
Endergonic - Requires energy, uphill reaction, energy requiring
Exergonic - Releases energy, downhill reaction, energy release
Activation energy
Denature - Active site has changed and thus the enzyme becomes useless
Optimum temperature
Saturation point
Substrate concentration
Cofactors
Metal ions
Key Knowledge:
Enzymes and coenzymes are catalysts that assist with photosynthesis and cellular respiration
Factors that impact on enzyme function
Enzymes are proteins made from amino acids
Reusable
Speeding up chemical reactions by lowering the activation energy
Usually have the suffix ‘ase’ or ‘in’ (Eg. sudcrase, lipase, trypsin, pepsin)
Names can identify the substrate
Sucrase catalyses sucrose
Lipase catalyses lipids
Describes what the enzyme does or what fits in it
Substrates are substances that ‘fit’ into the active site of an enzyme.
Bonds are formed between the substrate, energy → water released and peptide
Active site is the region where only specific substrates are able to bind and undergo a chemical reaction
Substrate is the substance that can bind to the active site in an enzyme,
Enzyme specificity is based on the specific shape of the active site
When an enzyme binds to a substrate its called an enzyme substrate complex
Becomes enzyme product complex after catalysis
Theories:
Substrate and enzyme are directly complementary
Lock and Key Model
Perfect fits designed for each other
The active site and substrate are complementary
Induced Fit, the enzyme conforms for the substrate
The active site becomes complementary
Enzymes ARE proteins and are made of amino acids
Biological catalysts → speeds up processes
Not permanently changed in processes
Reusable
NOT reactants → written on the arrow in reaction equations
Catabolic Reactions
Large molecules to small molecules
Breaks down
Exergonic, releases energy
Needs water
Anabolic Reactions
Small molecules to large molecules
Builds
Requires intake of energy
Endergonic
Produces water
Activation energy:
Energy needed for any chemical reaction, any reaction will occur with enough energy. Enzymes work by lowering the required activation energy.
Factors affecting enzyme activity:
Temperature
Heat energy means more collisions between enzymes and substrates however enzymes denature so the rate of reaction falls
Optimum temperature is 37.5 degrees celsius
When there is low temperatures there is less kinetic energy
The enzyme is NOT denatured at low temperatures but it IS denatured at high temperatures
Low kinetic energy low molecule collisions less enzymes substrate complexes forming etc.
pH
pHs have an optimum pH for enzymes but enzymes can denature on both sides of pH, if it’s too low or too high.
Works only within a small pH range
pH disruption generally results in complete loss of activity
Identical parabola
Substrate concentration
Substrate graph optimum is the point of saturation
Point of saturation means all active sites have been occupied and the enzymes must produce at a stable reaction rate.
More enzymes = Increasing reaction rate
Increasing then plateau
Enzyme concentration
Rate of reaction can increase as long as there is enough substrate
If there is an overabundance of enzymes and not enough substrate no reaction will occur
Constant line
Cofactors & Coenzymes
Cofactors can either be metal ions or coenzymes
Metal ions bridge enzymes + substrate together, combined with the catalyst
Coenzymes, non-proteins, organic complex
makes the substrate fit better
All coenzymes are cofactors not all cofactors are coenzymes
Activate the enzyme
Inhibitors
Competitive and noncompetitive inhibitor
Competitive is COMPETING with the active site and binding to the active site
Non competitive inhibitors are not trying to bind to the active site, they bind to the enzyme changing the active site inhibiting the substrate from binding
More substrates can overcome competitive inhibition but cannot overcome noncompetitive inhibitors
Competitive Inhibitors are TEMPORARY and REVERSIBLE, because they can eventually leave the enzyme
Allosteric = not the active site
Non competitive inhibitors force the enzyme to change PERMANENTLY and are IRREVERSIBLE
Stirring/Agitation
Increases collision and increases reaction rate
Increases the substrate and enzyme collision
Feedback inhibition
Metabolic pathway
Dephosphorylation
Phosphorylation
Large molecules need to be broken down slowly over multiple stages
The last product made in a process can be an inhibitor for enzyme one which stops too much product being made.
Feedback inhibition
Metabolic pathway
Each pathway requires a specific enzyme
The pathway is stopped by feedback inhibition
If the gene coding for an enzyme is messed up then the lack of feedback inhibition will cause a build up of a certain substrate and there will be no product
Exergonic
Dephosphorylation - Losing a phosphate
Phosphorylation - gaining a phosphate
ATP
ADP
Phosphate
Metabolic Reactions
Mechanic work
Nucleotide
Hydrolysis
Hydrolyse
Loaded - Fully energised (ATP)
Unloaded - Not fully energised (ADP)
Inorganic phosphate (Pi)
Organic - molecule with carbon
Inorganic - molecule without carbon
Cells need energy to make muscles work, carry out chemical reactions, the growth and repair of cells, making larger molecules, maintaining body temperature
Enzymes are important in cellular respiration
Mechanical reactions, metabolic reactions
Adenine Triphosphate
1 Adenine (N-Base) + 1 Ribose Sugar + 3 Phosphates
Linked by hydrogen bonds
ATP breaks down and releases a lot of energy and becomes ADP and phosphate
ATP → ADP → ATP (reversible)
ATP is a nucleotide
Hydrolysis - To split ATP
Water is needed to split ATP into ADP
ATP is a loaded molecule
Enzyme used to catalyse ATP is called ATPase
Reaction type is Exergonic, Hydrolysis, Catabolic, Dephosphorylation
ADP is an unloaded molecule
ATP can act as a coenzyme and assist other enzymes
It is an energy carrying molecule
Provides enough energy to support reactions through the breaking of phosphate bonds.
ATP’s third phosphate is weakly bonded and has high energy
ATP is synthesised through cellular respiration
ATP synthase is enzyme to create ATP from ADP
Located in the mitochondria’s membrane and in chloroplasts
Found where ATP is needed to be made ie: mitochondria, chloroplast etc.
Dehydration/Condensation reaction = water being produced (ADP→ATP)
Endergonic, Anabolic
OTHER COENZYMEs:
Coenzymes are not specifically for substrates they are carriers to the reaction products
Coenzymes are regenerated to be reused
H+ (electrons)
NAD+ (+) H+ (+) 2e- (Unloaded → NADH (Reduction) (Loaded)
The other way round is Oxidation
FAD (Unloaded) → FADH2 (Loaded)
NADP→ NADPH
Workbook 20-21
CELLULAR RESPIRATION
Reactants
Cellular resp
Glycolysis
Krebs cycle - Takes place in the mitochondrial matrix
ATP yield (NOT NUMBER IN TEXTBOOK)
Electron transport chain - Set of reactions in the mitochondria
inputs/outputs
Locations of glycolysis
Coenzymes
Pyruvate/Pyruvic Acid (3 Carbon molecule, break down of glucose)
Intermediate reaction
Cristae - Mitochondrial inner membrane, most ATP is made
Mitochondrial matrix is the fluid in the mitochondria
Glucose + Oxygen ⇒ 30-32 ATP + (Water) + (Carbon Dioxide)
CO2 and Water are byproducts the intention is for ATP
1 glucose makes 30-32 Total ATP which is too much energy to be produced at once
Glucose | Oxygen | Carbon Dioxide | Water | ATP | |
Animal | Ingested | Inhaled | Exhaled | Output | Energy currency to maintain life |
Plants | PHS product | PHS product | PHS input | PHS input |
All cells respire
Respiration is a set of metabolic reactions
Purpose: To convert glucose to ATP
Mitochondria is the powerhouse of the cell
Wherever ATP is being made there is ATPsynthase
When glucose enters a cell it breaks down with glycolysis, taking place in the cytoplasm
The glucose breaks into Pyruvate x 2which breaks down into CO2 x 2 and Acetyl CoA x 2
Pyruvate to CO2 and Acetyl CoA is an intermediate reaction
Acetyl CoA then goes to the mitochondria’s matrix which is a fluid where the Krebs cycle occurs
When glucose → pyruvate → acetyl CoA is made Hydrogen ions are released which would disrupt the pH of the cell, NAD + FAD pick up the hydrogen ions and become NADH or FADH2 which are reduced, loaded molecules and are used to transport to the cristae
NAD and FAD are electron carriers/coenzymes
NAD is an important coenzyme that is used to activate lactic dehydrogenase enzyme
Intermediate reaction = link reaction = transition reaction
Glycolysis → Intermediate reaction → Krebs cycle → Chemiosmosis
Glycolysis produces 2 ATP and Krebs produces 2 ATP and Electron transport chain produces 26-28 ATP so in TOTAL 30-32
Glycolysis - Cytosol
Matrix - Kreb cycle
Cristae - ETC
Mitochondria has;
Two membranes, Outer and Inner (Cristae)
Shaped for maximum efficiency
Requires 2ATP and ATPase
Breaks down into Pyruvate or Pyruvic acid
NAD+ come and collect the hydrogen ions
2 ATP is produced from both ‘breakdowns’
Total of 4 ATP produced but net 2ATP because there is an investment of ATP
Input 2 acetyl coa 2 Adp 6 NAD+ 2 FAD+
Output 4CO2 2 ATP 6 NADH 2 FADH2
Only 2ATP are produced
Oxygen is NOT involved
More electrons are picked up by NAD and FAD
All co2s have been released
Made in the MATRIX
NADH is oxidised and GIVE UP its hydrogen ions
Made in the Inner mitochondrial membrane or the cristae
The hydrogen ions cross the cristae membrane
All the hydrogen ions then go through ATP synthase and make ADP into ATP
26-28 ATP
Facilitated diffusion through ATP synthase
Electron transport chain = oxidation phosphorylation
H+ cross membrane → proton gradient (concentration) increases outside membrane → H+ is then facilitated diffusion-ed through ATP synthase → ATP made
Oxygen is the final electron acceptor, water is made as oxygen + H+ + electrons
10 mill ATP is produced per second by one cell
Fermentation
Alcohol Fermentation
Lactic Acid Fermentation
Anaerobic Respiration
Muscle Fatigue
O2 Debt
Ethanol
Lactic Acid
When oxygen is not present in the cells, the process of fermentation occurs
Glycolysis → Fermentation
NADH returns the H ion and the pyruvate becomes lactate or ethanol
Reversible once oxygen becomes available
Lactic acid in animals, bacteria and some fungal cells
Alcohol/Ethanol produced in plants and yeast
2(C3H6O3)
Lactic acid is poisonous to the body and builds up creating muscle cramps, it is broken up by oxygen
Ethanol is 2C2H5OH created by plant and yeast fermentation
Lactate dehydrogenase and Alcohol dehydrogenase
Glucose → ethanol + carbon dioxide + energy (2ATP
Glucose → Lactic acid + energy (2ATP)
When animals run out of O2, pyruvate becomes lactic acid creating muscle fatigue and O2 debt
Provides rapid bursts of ATP in muscle cells
But is incredibly toxic
Oxygen concentration
Temperature
Glucose availability
Hydration, light, age, activity level
Respirometer is a device that determines an organisms respiration rate by measuring the rate of exchange of O2 and CO2
Living specimens enclosed in sealed container
Pressure changes affect the manometer
Increasing Co2 levels or decreasing oxygen levels → Increases respiratory rate
**RATE OVER TIME (over time must be included)
Temperature denatures enzymes therefore pathways cannot continue
Oxygen measured by the amount of CO2 produced
Increase O2 —> increase resp. Rate until point of saturation
Anaerobic respiration is measured by lactic acid of alcohol produced, aerobic respiration measured by Co2 levels
Glucose and oxygen are substrates
Glucose conc increases
Biofuels - reducing the amount of greenhouse gases by being renewable and recyclable fuel made from recently living organisms such as plants and algae
Fossil Fuel
Biomass
Fermentation
Renewable
Burning biofuels releases carbon dioxide but they are carbon neutral because they cancel out the amount they release by holding that amount of carbon dioxide when being grown.
Needs to be easy to make, transport and able to mix with fossil fuels
Places to grow this.
Processes requiring energy
Feedstock | Advantage | Disadvantage |
Corn, Rapeseed |
|
|
Waste cooking oil, Lignocellulosic feedstock |
|
|
Microalgae |
| Low lipid content Contamination problem |
Engineered microalgae |
|
|
Two main types of biofuels, bioethanol and biodiesel
Bioethanol/Ethanol
Fermenting sugarcane /starchy plant materials
Blended with petrol
Biodiesel
Made from lipids/fatty acids
Inputs, outputs
Roles of enzymes and coenzymes in facilitation
Locations of light dependent and light independent stages
Rate of photosynthesis and factors that affect rate
Factors: light, water, temperature, carbon dioxide
Photosynthesis
Light dependent
Light independent
Autotrophs, producers
Heterotrophs
Palisade layer - generally the more photosynthetic cells
Epidermal layer
Spongy layer
Mesophyll - all the in between bits (palisade and spongey)
Bundle Sheath cells
Stomata/Stomate
Guard Cells around the stomata - close at night, open day which allows for the diffusion of Co2
Oxidation
Reduction
Xylem tubes - Carry water
Phloem tubes - Carry food
Veins
Granum is the pile of discs or Thylakoids
Thylakoid, the membrane of the discs in a chloroplast, is the disc
Stroma
Inner Membrane
Outer Membrane
NADP and NADPH
Calvin cycle
Limiting Factor
G3P
PG3 - Phosphoglycerate
Glucose
C3
C4
CAM
Turgid
Flaccid
Sunlight is needed because it is the initial energy source
Plants make glucose during daylight, as long as there is sunlight
They require glucose because they are autotrophs, they make their own food
Water is diffused by osmosis
Inputs: Water + Carbon Dioxide
Outputs: Sugar + Oxygen
Input/Output and Light | Location | How |
Carbon Dioxide | Stomata | Gas |
Water | Roots | Osmosis |
Light + Chlorophyll | Leaves, Chlorophyll | Chlorophyll which traps light |
Sugar/Glucose | Stored as Starch and used in mitochondria in leaves and phloem | |
Oxygen |
Include light and chlorophyll on the →
Cuticle protects plant
Epidermal Layer is made of epidermal cells
Palisade Layer, made of palisade cells
Sponge layer, made of sponge cells and air
Stomata used for co2 diffusion and water transpired out
Photosynthesis takes place in the chloroplast
Chlorophyll, containing ATP synthase
6CO2 + 6H2O → C6H12O6 + O2
6CO2 + 12H2O→ C6H12O6 + 6O2 + 6H2O
Reduction is gain of electrons
Oxidation is loss of electrons
H2O is oxidised, CO2 is reduced
NADP comes and picks up the hydrogen ions released from the reaction and split of H2O and comes and picks up the ions and becomes NADPH
Oxidation reaction = water being oxidised to oxygen through NADP
Reduction reaction = Carbon Dioxide is reduced to glucose
Glucose is made for cellular respiration
Xylem tubes carry water from roots to leaves and parts of the plant through veins
Phloem tubes carry food to all parts of the plant body for all cells to respire
Made up of granums which are discs filled with chlorophyll and membrane known as Thylakoids
Water is trapped in the Thylakoids and is split releasing Oxygen to be diffused out of the cell, some may be used in respiration but the rest will diffuse into the atmosphere
NADP comes and collects the hydrogen from the split H20 becoming NADPH
The H+ is used to synthesise glucose
Light dependent stage, splitting of H20 takes place in the Thylakoid
The Calvin Cyle or the synthesis of glucose and the reduction of Co2 doesn’t require light and is the light independent stage
ADP + Pi becomes 18ATP
Photosynthesis is photo, then synthesis
Photo - light dependent
Thylakoid/Granum
Chlorophyll traps the light to split water and produces 18 ATP and O2
Synthesis - light independent
Stroma
Calvin cycle
Using 18 ATP from light dependent reaction
Photosystems 1 and 2 are photosynthetic pigments that absorb light energy
Calvin cycle: CO2 + NADPH + 18ATP → NADP + 18ADP + GLUCOSE
Occurs in the stroma and doesn’t require light but depends on products of light dependent stage
Uses ATP and NADPH to synthesise glucose
Endergonic and Anabolic
NADPH → NADP
Enzyme rubisco is required
It fixes RUBP (sugar with 5 Carbon) to CO2 and turns it into a 6 Carbon molecule
Which becomes 2 x G3P
One G3P will leave as an output of SUGAR
The other will continue in the cycle and become RUBP
A higher concentration of CO2 means more reaction
Calvin cycle runs 6 times before producing glucose
Rubisco is an enzyme for Carbon Dioxide but when there is a high concentration of Oxygen, oxygen will bind to Rubisco instead.
External Factors include:
Light
Temperature
CO2
Water
Internal factors
Chlorophyll - affects the amount of light being absorbed - is a limiting factor
Limiting factors are factors that limit the reaction such as limited by temperature being too low or temperature being too high etc.
Photorespiration
When there is no CO2 entering due to closed stomata and increase in O2 concentration then photorespiration occurs
Rubisco picks up O2, high temp, stomata closed, when oxygen is more concentrated than co2 and creates CO2 by binding O2 to Rubisco
Stomata/Stomate - openings guarded by the guard cells
Guard cells turgid/Stoma Open
Turgid = Full
Cells flaccid/Stoma closed
Guard cells use osmosis to open and close by the concentration of sugars
C3 plants have thin bundle sheath cells
C3 make up most plants
Occurs only in mesophyll cells
C4
Pep Carboxylase ( PEPCase) always binds to CO2 to produce C4 Acid Oxaloacetate
More efficient than Rubisco because it will always collect Co2
PepCase makes Malate and CO2 in the Mesophyll cells from Co2 and PEP Carboxylase
Co2 Increases, Rubisco picks up Co2 to fix to RUBP and continue the calvin Cycle
Occurs in Bundle sheath then Mesophyll
C4 | C3 |
2 Carbon Fixations | 1 Carbon Fixation |
Photosynthesis | Photorespiration and Photosynthesis |
Adapted to low light and low water, more efficient | Not as good in hot weather, closes stomata in hot weather |
Thick Bundle Sheath Cell | Thin bundle sheath cell |
1st product is Oxaloacetate (4p) | 1st product is PGA (3p) |
CAM
Closes stomata during the hottest part of the day
Traps Co2 at night time
Making sugar with trapped CO2
Similar to C4 plants uses PEP carboxylase to malate fixation of CO2 and then to the Calvin Cycle
Occurs in Mesophyll all the time however the different processes are at night vs day
Comparing | C3 | C4 | CAM |
# of CO2 Fixation reactions | 1 | 2 | 2 |
1st stable product from Co2 Fixation | PGA (3C) | Oxaloacetate (4C) (Malate) | Malate (4C) |
Where Calvin Cycle occurs | Mesophyll cells | Bundle Sheath Cells | Vacuole & Mesophyll cells |
Photosynthesis and/or Photorespiration | Photosynthesis and Photorespiration | Photosynthesis | Photosynthesis |
Efficiency of CO2 Fixing | Poor | Good | Good |
When Stomata open | Day | Day | Night |
Best adaptation | Moderate cool and wet | Hot and sunny | Very hot and Dry |
Proteins
Hormones
Structural proteins
Biomacromolecules
Carbohydrates
Lipids
Nucleic Acid
Monomers
Polymers
Polymerisation - Making polymers from monomers, a reaction that combines monomers to make polymers
Oligomers
Alpha helix
Beta Pleated sheet
Polypeptide
Peptide bond
Primary, secondary, tertiary, quaternary protein structures
Globular Protein
Filament
Protofilament
Fibrous Protein
Proteomics - Study of proteome
Proteome - the complete array of proteins produced by a single cell/organism in a particular environment is called the proteome of the cell/organism
Purines - A & G
Pyrimidines - T & C
Chromatins
Messenger RNA
Transport RNA
Ribosomal RNA
Transcription - Occurs in nucleus, copying Dna code onto mRNA
Translation - Translating mRNA (Decoding)
Proteins are used for everything such as contraction, reception, hormones, protection, transport, storage, enzymes, structural, identification, signal etc.
They are coded from Amino acids which are building blocks
Contain C, O, H, N & S
Proteins are building blocks. They have a wide range of functions
Monomers are the basic building block
Amino acids are the monomers for proteins
Nucleotides are monomers for nucleic acids
Monosaccharides are monomers for carbohydrates
All amino acids have
An amino group
A carboxyl group 0=C-OH
A unique side chain (often depicted as R) H-C-R
Amino Acid 1 + Amino Acid 2 → Dipeptide + Water
Condensation/Dehydration reaction because production/loss of water
Amino acids combine and peptide bonds form from amino acids to become a polypeptide.
The covalent bond between the amino acids is the peptide bond
Polypeptide chains can be broken down via hydrolysis reactions which splits the chain.
4 levels of protein structure
Primary protein structure - Linear sequence of a chain of amino acids
Secondary protein structure - an alpha helix + a beta-pleated sheet which is the folding of polypeptide chains into helices or sheets
Tertiary protein structure - 3D folding pattern of a protein due to side chain interactions - Globular protein, very specific shape being formed
Some proteins only continue to Tertiary structure but some combine into more than one amino acid chain
Quaternary Protein structure - Protein consisting of more than one amino acid chain
Linear sequence will provide information on how the protein folds, function or no function, evolutionary relatedness between species.
Hydrogen bonds hold alpha helix, peptide bonds form between amino acids
SECONDARY PROTEIN STRUCTURE
Alpha helix
Beta Pleated sheets
Random Coils
TERTIARY STRUCTURE
Total irregular folding and bending of chain
Causes amino acids to become close
Function depends on shape
3D protein
Disulphide bonds only existing in tertiary structure
Hydrogen bonds
QUATERNARY
Fibrous or globular
Some are conjugate containing inorganic compounds
Eg. Haemoglobin
The two types of nucleic acids
DNA + RNA
Made of a sugar, phosphate and nitrogenous base (ATGC or AGCU)
RNA is single stranded
DNA is double stranded and also has no oxygen - Antiparallel double stranded helix
AT are double bonded
GC are triple bonded
The phosphate makes DNA negatively charged
The units of DNA inside the nucleus are Chromatins which when needed to split become Chromosomes.
5’ and 3’ Antiparallel structure (‘ = prime)
Three types of RNA
Messenger RNA - Carries instructions for polypeptide synthesis
Ribosome - Structural subunits of the ribosome
Transfer RNA - Carries amino acids to ribosome
DNA are the instructions the RNA are the messengers
RNA copies a strand of DNA and then this is used to go to the ribosome which codes 3 bases at a time using amino acids taken from food and the breakdown of proteins
DNA is in the mitochondria, chloroplast and some in the mitochondria only 1 type
RNA in the nucleus and cytoplasm has at least 3 types
Made in 3s in the ribosome
Transcription occurs in the nucleus - the copying of DNA onto mRNA
Translation in the ribosome - decoding of mRNA into the ribosome
Gene Expression = Protein Synthesis
Transcription
Translation
Ribosome
Protein Synthesis
Sense/Antisense strand
Template/Non-template strand
Amino acids
Non-Coding/Coding
Pre-RNA
Elongation
Binding
Initiation
Promoter region
Methylated Capping
Introns
Exons
Gene sequence
Exon juggling
DNA codes the instruction for protein synthesis but
Ribosome is the site of protein synthesis which is in the endoplasmic reticulum
DNA cannot leave nucleus but Ribosomes cannot enter the nucleus
A copy of DNA is made as RNA through transcription
At the ribosomes, the copy is translated by tRNA and the necessary amino acids will be produced
Template strand/Antisense Strand/Non-coding strand
Sense/Non-template/Coding strand
Gene sequence = correct order of nucelotides
Steps of Transcription
1.Initiation: Initiation factors (proteins) bind to DNA stand to switch on the gene
2. Binding: RNA polymerase binds to the promoter region of the template strand
3. Elongation: RNA polymerase moves along the template stand, preliminary to RNA
Methylated Capping and poly-deniylation tail for Transcription
Introns and Exons
Base pairs are read by tRNA three base pairs at a time
tRNA structure consists of a structure of amino acid and anti-codon
TRANSLATION STEPS
mRNA attaches to a ribosome
tRNA (anticodon) attaches to mRNA (codon) (base pairing)
A specific amino acid is detatched to form either a polypetide or a peptide bond with an amino acid
Stop Codons are codons that literally stop the process of forming the polypeptide chain etc.
The stop codons are UAA, UAG, UGA
All living organisms have the same amino acids
There are more than one codon for each amino acid to allow for error
Gene Regulation - when a gene is switched on
Structural gene - gene that encodes for a specific protein
Regulatory gene - A gene that encodes for protein that regulates the structural gene ie activators or repressors to switch the genes on and off, codes for regulatory proteins
Activator proteins - turns genes on to start transcription
Repressor proteins - turns the gene off to prevent transcription
Promoter region - where RNA polymerase binds
Operator region - where regulatory proteins binds
Operons - only occurring in prokaryotic cells, a functional unit of transcription that regulates gene expression in bacteria
TRP Operon
Tryptophan
Anti-termination loops
Termination Loop/Attenuator stem loop
Hairpin loops
Attenuation
Attenuator
Lead mRNA
trpL
Leader region
Genes need to be regulated because not all cells can make every single protein all the time so certain cells are made to produce certain proteins and certain genes are on or off
The controlling of gene expression
Structural gene
Regulatory gene
The regulatory protein (activators or repressors) binds to the Operator region
Prokaryotic Gene Regulation
Does not have introns or exons or gene capping
Can combine multiple structural genes to create an operon
Occurs in the cytoplasm
TRP OPERON
The TRP operon is an operon in bacteria
Made by bacteria by the TRP Operon
Tryptophan - Amino Acid
The TRP operon codes for enzymes that catalyse the creation of Tryptophan
When TRP is high - Operon turns on
When TRP is low - Operon turns off
The TRP can regulate the levels
Only in prokaryotes
An Operon is a cluster of genes under the control of a single promoter
Transcribed and then Translated
When the operon needs to lower the amount of trp enzymes the regulatory region will make regulatory proteins that switch structural genes on and off
Tryptophan levels high → Tryptophan binds to the Repressor, and RNA polymerase cannot bind
ATTENUATION
Alternative method of reducing the expression of the trp Operon in prokaryotic cells
It relied on the capacity for prokaryotes to be both transcribing and translating SIMULTANEOUSLY
It prevents transcription from being completed
It can occur when tryptophan levels are high as a backup to terminate transcription when the repressor detaches from the operator
Stops further synthesis of the creation of tryptophan
In the trpL (lead protein)
The leader region of the operon is the trpL leader + Attenuator
trpL codes for the leader mRNA
Four regions that can form base pairs to form 3 hairpin loops
When there are low levels of tryptophan, the antiterminator loop occurs and blocks the formation of the termination loop (attenuator) it pauses the protein synthesis
The attenuator is like a stop codon and will stop transcription, the termination loop will cause the ribosome to stop translation
there are trp codons on the trpL when the tryptophan levels are high and it will pause the ribosome at the stop codons which covers the 2 and stops 2 and 3 from binding and then allowing the termination loop
When the 3rd loop forms stuff don't work
Vesicles - membrane bound packages
Transport Vesicles
Secretion Vesicles
Synthesis
Exocytosis - leaving the cell
Lysosomes - vesicles with digestive enzymes
Golgi apparatus
Smooth ER
Rough ER
Proteins are transported in vesicles from the Rough ER to the golgi apparatus and are then transported to be secreted from the cell membrane or used around the cell
Smooth ER for hormones and lipids
Rough ER usually for proteins
REVISION QUESTIONS:
Tryp Operon
What is the role of the trp operon in regulating the synthesis of tryptophan?
The TRP operon regulates the production of the enzymes for the biosynthesis of tryptophan.
What type of gene regulation is involved in the regulation of the trp operon?
Negative regulation.
How does the presence of tryptophan affect the expression of the trp operon?
HIgh levels of TRP make the trp repressor fall off the operator region by binding to the repressor, in low levels, the repressor falls off. If the repressor falls off while trp levels are high then attenuation is used.
What is the role of the trp repressor in regulating the trp operon?
It prevents RNA polymerase from transcribing the leader genes and the structure genes so that the enzymes for the biosynthesis of tryptophan cannot occur.
How does attenuation regulate the expression of the trp operon?
It acts as an emergency in case the repressor is not bound to the operator region of the operon when tryptophan levels are high. By not pausing RNA polymerase when it reaches region 1 and pausing at region 2 it allows regions 3 and 4 to bind creating a hairpin loop that forces the ribosome to fall off.
Gene Regulation
What is gene regulation and why is it important for an organism?
Gene regulation is the control of the production of proteins based on gene expression and it is important for specialised cell functions and proper cellular development.
What are the different types of gene regulation mechanisms?
Negative and positive regulation, splicing, attenuation, feedback inhibition, transcriptional regulation, translational regulation
What is the role of transcription factors in gene regulation?
How do epigenetic modifications affect gene expression?
What is the difference between positive and negative gene regulation?
Protein Packaging
What is protein packaging and why is it important?
What is the structure of a nucleosome and how does it package DNA?
How are histones involved in protein packaging?
What is chromatin and how does it affect gene expression?
How does the packaging of DNA affect the accessibility of genes for transcription?
Protein Synthesis
What is protein synthesis and what are the two main stages involved in it?
What is the role of messenger RNA (mRNA) in protein synthesis?
What is the function of ribosomes in protein synthesis?
How is the genetic code translated into a specific amino acid sequence?
What is the role of transfer RNA (tRNA) in protein synthesis?
Nucleic Acids
What are nucleic acids and what is their function in cells?
What is the structure of a nucleotide and how are nucleotides joined together to form a nucleic acid?
What is the difference between DNA and RNA?
What is the function of DNA in cells?
What is the central dogma of molecular biology and how do nucleic acids play a role in it?
Proteins
What are proteins and what is their function in cells?
What is the structure of an amino acid and how are amino acids joined together to form a protein?
What is the difference between a primary, secondary, tertiary, and quaternary structure of a protein?
What is denaturation and how does it affect the function of a protein?
What is the role of chaperone proteins in protein folding and quality control?
Biotechnology - use of an organism or organism component to make a product or process (ie: COVID vaccine)
DNA technology - sequencing, analysis and cut & paste of DNA
Meiosis
Genetic Recombination - Exchange of info between 2 DNA segments
rDNA - recombinant DNA technology
Vectors
Plasmid
Cloning Factor
Amplified Gene
Amplified Protein
Recognition sequence - identifies if something needs to be cut
Restriction enzymes
Reverse transcriptase
Ligase
Polymerase
Sticky ends
Blunt ends
Ligation
VCAA Knowledge: The use of enzymes to manipulate DNA including polymerase to synthesise DNA
Majority of Biotechnology relies on DNA manipulation
DNA technology is the sequencing, analysis and cut & paste of DNA sequences
Genetic Recombination which uses or occurs in meiosis and exchanges info between 2 DNA segments (like homologous chromosomes)
Occurs b/w same species
Makes a recombinant chromatid (or non-recombinant)
DNA manipulation literally manipulates genomes/genes to introduce or take away specific objectives.
rDNA is recombinant DNA technology
Genetic engineering, recombinant biotechnology, DNA manipulation
Gene technology includes manipulation and analysis of DNA
DNA manipulation alters DNA by adding or editing DNA
Bacteria is often used as vectors
To reproduce an edited gene a cloning vector is needed such as a plasmid from Bacteria to create a recombinant plasmid
Protein or Gene can be amplified
In order to → Tools used
cut DNA: Restriction enzymes (Endonuclease)
Stick DNA fragments: DNA ligase
Copy of DNA: Polymerase
Make multiple copies of DNA: Polymerase Chain reaction (PCR)
Separate DNA fragments: Electrophoresis
Edit Genes: CRISPR
To synthesise DNA, replicate or repair
Copy DNA and make copies
Example: Taq polymerase used in PCR
DNA Polymerase creates DNA by asembling 749 (approx) nucelotides per second
Synthesis from 5’ to 3’
Taq Polymerase is a DNA polymerase used on PCR (chain reaction)
Joins different pieces of DNA
Used to make recombinant DNA
Recombines the DNA
Ligase closes the ‘nicks’ in the phosphodiester bonds + close the gaps and seals the newly transferred dna segment
Same Restriction enzymes cut gene of interest and plasmid /cloning vector
Plasmid and gene fragments connect and anneal
Ligase used to anneal the recombinant plasmid and seal it
Ligation
Cuts DNA and creates fragments
Examples: BamH1 (Bam = bacteria, H = Strain, 1 = Order of discovery)
EcoR1 and Taq1
EcoR1 - GAATTC (recognition sequence) will cut between the G and A so long as there is an AATTC following.
Sticky ends and blunt ends when one is overhanging or when there is a clean cut
Obtaining DNA from RNA
Reversing the transcription
PCR
Heating and Cooling Cycles
Amplify
DNA segment
Target DNA sequence
Primers - complementary segments that will attach to the 3’ prime end of a chain, extend the DNA through complementary base pairing
Buffer Mix - Maintains pH
Taq polymerase -
PCR is a technique to amplify or produce copies of a DNA segment
Using heating and cooling cycles
Replication at an exponential Rate
3 Steps
Denature
Separating the hydrogen bonds (94-95o)
Anneal
Primers bind to template (50-56o)
‘Annealing of the primer when it ‘cools’
Extension
Increases in temperature to 72
Synthesising a new strand
PCR can be used for;
consumer genomics
Food and agriculture
Forensic science
Genetic research
Medicine
Phylogenetics
Environmental biology
State purpose of PCR
To replicate specific DNA segments
Identify components
Primer
Nucelotide
Heat
Buffer
DNA sample
PCR Tube
Draw Steps to illustrate basic process
Heat stuff up and break hydrogen bonds
Anneal stuff with primers at the 3’ ends
Synthesise new strand
Identify Two applications
Gel Electrophoresis - used for sorting dna fragments
Agarose Gel - Jelly (containing buffer solution to maintain pH)
Buffer
Gel Matrix
Cathode
Standard - Kind of like a ruler used to compare dna sizes
Loading Dye
Ethidium Bromide - Fluorescent dye, binds to dna, mutagen
DNA fragments put into gel, and has positive and negative charges on each side, DNA is negative charged cos Phosphates so is drawn to positive charge.
Gel/matrix traps larger fragments and therefore separates dna fragments by size
Separate mixtures
Calculate size
Components
Electric current
Agarose Gel - traps dna allowing small fragments to move further down
Loading Dye
Standar - to calculate sizes
Ethidium Bromide - Fluorescent dye, binds to DNA
Buffer solution
Combs - to create wells to load DNA
Positive and negative terminal
STEPS
DNA fragments loaded in neg side
Electric current
Dna fragments migrate towards pos side
Small fragments move easily and travel far distance/larger fragments are trapped and travel less
Unit, Size (bp or Base Pairs) , Quantity (ng)
Sometimes known as a ladder (the standard)
DNA profile
Polymorphic regions - regions that have more than one change
Polymorphisms - differences in these region
Probable Origin
Microsatellites - they are STRS
STRs/Short tandem repeats
Non-coding DNA
Repeating code
Genetic loci (location)
Heterozygous genotype
Combined DNA index System or CODIS
Can be used to identify probable origin
Reveal family relationships
Identify Victims in disasters
Paternity Tests
Find Evolutionary relationships between species
Small sections in DNA vary, everything else is identical, these small sections identify individuals
Everyone inherits unique combination of polymorphisms
Process: Collecting DNA then cutting with endonuclease and then using gel electrophoresis to separate and identify.
STRS are regions of non coding DNA that contain repeats of the same nucleotide sequence
Short → 1-9 base pairs long
Tandem → Repeating Code
STRS are found in different places/genetic loci
Microsatellites (1-9bp), Minisatellites (10-100bp), Macrosatellites (>100bp)
Minisatellites for fingerprinting
STRS/Microsatellites for DNA profile
STR-XX (x,y)
If it’s different length, then its heterozygous
If same length, homozygous
ETHIC ISSUES
Limits of testing
Advantages | Disadvantages |
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Plasmids
Bacterial Chromosome
Recombinant
Colony
Vector
Antibiotic plate
Transgenic organisms (TGO)
Genetically modified organisms (GMO)
Cloning Vector
Gene of interest
Foreign gene
Foreign cell
Bacteria have both Plasmid DNA and Bacterial DNA
Plasmids are used as cloning vectors in rDNA technology because they can
Replicate
transfer genes from one cell to another.
Bacterial transformation is when foreign DNA is transferred
Bacteria are able to take up new DNA very quickly, coming from other cells
Changing the Genome/DNA transfer = Bacterial Transformation
pGLO is a plasmid from a jellyfish that glows
They can put the plasmid in via Heat shock and Electric shock
Heatshock: By putting bacteria into very cold solution, plasmids are added, then in hot water the plasmid and bacteria combine, then in ice bath again.
pGLO, ampicillan etc. has an antibiotic resistance gene
Some of the bacteria will take up the plasmid but some will not
To find the recombinant plasmid, they use an antibiotic plate and the antibiotic resistant plasmid will form a colony and show that the plasmid has affected it.
A vector is a molecule used as a vehicle to carry genes of interest to foreign cell.
Bacterial plasmids are commonly used because of self replication etc.
These plasmids can be modified for specific usage.
PLASMID VECTORS
Multiple Cloning site - contains numerous recognition sites to allow gene insertion
Promoter - Initiates transcription
Origin of replication - for plasmid synthesis
Antibiotic resistance - selects modified cells
Reporter gene - makes products that attach to the protein to enable detection
STEPS IN PLASMID CLONING
Restriction enzymes to digest DNA sample and DNA plasmid
DNA ligase to seal
Transformation of recombinant plasmid into bacteria
Agar plates with selection antibiotic resistance allow bacteria with the resistance and gene of interest is cloned.
ANTIBIOTIC PLATE
The antibiotic plate is used to see if the transformation has occurred, as the antibiotic plate will kill any bacteria that has not been transformed, and transformed bacteria will grow because they have the antibiotic resistance and therefore show what has and has not been transformed.
Can be used to increase gene pool
Humilin
Insulin
Cloning Vector
A Chain - 21 Amino acids
B Chain - 30 amino acids
cDNA - copy DNA
‘S’ - suppression not resistant
‘R’ - resistant, immune
Fusion protein - A protein made from a fusion gene which is created by joining parts of two different genes.
Has 51 amino acids
Is a hormone produced by beta cells
Lowers blood glucose
Has a quaternary protein structure consisting of 2 peptide chains held together by disulphide bonds between cysteines
Chain A - 21 amino acids long
Chain B - 30 amino acids long
A plasmid vector and insulin gene are isolated from E.coli then it is cut open by the same restriction enzyme.
1 bacteria is used to make Chain A and a different bacteria is used to make chain B so that we have complete control over when chain A and B are together and manufactured.
Insulin will only become functional when its taken from the two cells.
When DNA is transcribe into mRNA then reverse transcription occurs its a DNA (single), polymerase is needed to make it double stranded.
Synthesis of Chain A/B
Obtain copy of the insulin A gene that is double stranded without introns
It must not have introns because bacteria do not have introns and cannot function with such.
Double stranded to anneal to the double stranded plasmid.
cDNA is copy DNA
pBR322 has ampicilin resistance and TET resistance. It will be used to insert the insulin A gene which will make the CHAIN A protein.
pBR322 is the cloning plasmid.
When only one restriction enzyme is used, the sticky ends may stick back together, so two restriction enzymes are used. EcoRI and BamHI
Decrease the risk/chance of the plasmid coming back together.
Now a recombinant plasmid that has ‘Insulin A gene’ and loses its TET resistance
TET is interupted
WHen rplasmid mix with E.coli, some of the rplasmid will be transferred to E.coli to make them transformed, but not all will be transformed.
Rplasmid - AMP(r) + InsulinA gene + Interrupted TET gene
3 POSSIBLE OUTCOMES
Bacteria with NO pBR322
Untransformed
NO resistance to tetracycline or ampicillin
Bacteria WITH pBR322
Transformed bacteria by plasmid
Resistance to Tetracycline and ampicillin
Bacteria with Recombinant pBR322 containing the insulin Gene:
Transformed bacteria by rplasmid
ONLY resistant to ampicillin.
Method 2 uses Beta - galactosidase gene that codes for B-galactosidase enzyme which breaks down lactose and glucose and galactose
It’s used as a marker to identify the plasmids that have the insulin A/B gene, by changing colour
B-gal is to next to the Insulin A gene and goes into.
Gene expression of this plasmid
Beta galactose
Insulin A gene
Resistance to ampicillin
3 POSSIBLE OUTCOMES:
Untransformed bacteria
Did not take any plasmid
Not resistant to ampicilin
Transformed bacteria
Resistant to ampicillin
Insulin Gene
No Beta Galactosidase gene → No fusion protein made
Transformed bacteria
Resistant to ampicillin
Insulin gene
Beta galactosidase gene - Fusion protein made.
To identify which colonies have functional insulin gene (ie: Beta-galactosidase gene) bacteria will be grown on agar plate with X gal.
X-Gal detects the presence of Beta galactosidase and insulin gene by causing bacteria; colonies to have a blue or white colour.
BLUE colony: Beta galactosidase & Insulin Gene present
WHITE colony: NO Beta Galactosidase and Insulin gene.
Ethics
Bioethics
Moral principles
Human rights
Welfare of people
Informed Consent
Ethics: A system of moral principles, right v wrong
Bioethics: Moral principles specific to biological science
Beneficence - must not hurt others, maximising benefits and minimise harm
People welfare is prioritised over interests of science
Informed Consent
Holding healthcare institutions accountable and reviewing scientist works
Unethical things;
Plagiarism
False reports
Dishonesty
Breaches in integrity
Strict guidelines for evaluation, publication and follow up
Ethical Approaches to Bioethics:
Consequence bases
Places central importance on the consideration of the consequences of an action (the ends)
Aims to achieve the maximisation of positive results, with the minimisation of negative results.
The focus is on the eventual outcome as opposed to the process taken to reach it
What is the end result? Does it outweigh the negatives of the process?
Does the ends justify the means?
Duty/rule base
Duty of Care
concerned with how people act (the means) and the process taken to get to the result.
places central importance on the idea that people have a duty to act in a particular way,
•and/or that certain ethical rules must be followed, regardless of the consequences that may be produced.
Ie: it is not acceptable to cause immediate or temporary harm in the pursuit of a potential ‘greater good’
Virtues based
Is person-based rather than action-based.
Consideration is given to the virtue or moral character of the person carrying out the action.
Providing guidance about the characteristics and behaviours a good person would seek to achieve to then be able to act in the right way.
Things that are self-seeking are unethical
Use keywords: Outcome, good virtues, process, means, etc.
Ethical concepts are used when identifying bioethical issues and are used to inform ethical guidelines
When deciding the extent to which the outcome of something or the course of action is ethically applicable
INTEGRITY:
The commitment to searching for knowledge and understanding.
Honest reporting of results, sources, in ways that permit scrutiny and contribute to public knowledge and understanding
Regardless of favourable or unfavourable results
Must be transparent
JUSTICE
The moral obligation to ensure fair consideration of competing claims
No unfair burden on a particular group from an action. Fair distribution and access to the benefits of an action.
BENEFICENCE
Maximising benefits
Minimising risks and harms in taking a particular course of action or position.
‘Zero harm’
NON MALEFICENCE
Avoiding causation of harm
The harm must be less than the benefits of the courses of action and outcomes.
Harm/Risk can be a little as long as it is outweighed
RESPECT:
Consideration of the intrinsic value of all living things
Regarding welfare, liberty, autonomy, beliefs, perceptions. Customs. Culture
Considerations for agency
When living things have diminished capacity to make their own decisions ensuring they are protected and empowered.
Photosynthetic efficiencies
Crop yields
CRISPR-Cas9
Bacteriophage
gRNA - guide rNA
sgRNA - single guide RNA
CRISPR is used for Gene editing to ‘find and replace’
CRISPR Cas-9 is an Endonuclease complex that naturally exists in bacteria to edit virus DNA (bacteriophage)
Clustered Regularly Interspaced Short Palindromic Repeats
CRISPR has two important features:
Palindromic repeats : short palindromic repeats
The spacers are segments of viral DNA (bacteriophages) that allow the bacteria to recognise the same virus in the event of subsequent invasions.
CRISPR can:
Repair the damaged DNA by using a template with the correct sequence.
Silence a faulty gene: The DNA can ‘self repair’ when cas9 cuts it; however, mutations can occur to introduce the ‘STOP’ codon.
Replace faulty gene sequence with the correct sequence, by using cas9 to introduce enzymes that would replace faulty bases with the correct bases.
Increase transcription of a gene by deactivating cas9 and adding activators.
Complement proteins do three things,
1. Stimulate phagocytes to increase phagocytosis
2. Highlight/Tag invaders for phagocytosis
3. Cytolysis the lysis of a pathogen forming Membrane Attack Complex (splitting of a cell)
Cytokines are signalling molecules
Peptide, protein or glycoproteins
A chemical signal for cells to carrry out immunologic responses
Released in response to cell damage or to indicate presence of pathogen
Crucial in controlling growth and activity of other immune system cells and blood cells
Trigger cells to;
Proliferate (reproduce)
Induce inflammation
Promote antibody response
Activate macrophage
Interferons (IF) are cytokines, protein and signalling molecules
Act as a warning signal from infected cells to all other cells
Viral host cell is triggered to synthesise interferon which targets neighbouring cells
Allergens are harmless substances NOT harmful substances
ANTIGENS & MHC
Antigen = Antibody Generator
An antigen is a unique molecule (eg. protein) that triggers the production of antibodies and the immune system
Could be part of a microbe or foreign substance (does not have to be whole pathogen); bacteria, pollen, protein
Immune response may be
Antigen non-specific → innate immune response
Antigen specific → adaptive immune response
The presence of antigens classify blood type on Red Blood Cells
Antibodies?
Antigens are foreign to organisms and stimulate antibodies that are specific to that antigen
One antibody will generally bind to one antigen
Antigens are non-self/foreign molecules that the immune system recognises as enemies.
Cells are self or non-self, non-self cells have antigens.
Self Markers (MHC) label the body’s cells as a friend and are tolerated by the immune system.
MHC = Major Histocompatibility complex
If MHC is present, immune cells will not be activated to respond.
MHCs are a group of venus that code for proteins found on the surface of cells that help the immune cells distinguish between self and non-self cells
Two main types in vertebrates: Class I and Class II
Class I markers are found on body cells, somatic cells NOT red blood cells.
Class II markers are found on immune cells, macrophages, dendritic cells, monocytes, B cells, antigen presenting cells.
Class I: All nucleated cells eg. Body cells
Act to identify cells self antigens from non-self antigens
Class II: Primarily on professional APC [Antigen presenting cells]
These cells present Antigen fragments on their class markers to T lymphocytes and other immune cells. Immune cells will have both classes EXCEPT RED BLOOD CELLS.
Professional antigen-presenting cells - APC, identify antigens as non-self and process antigens
Present the antigen on a MHC marker, so that immune cells lymphocytes so that they can identify and destroy/respond to invaders.
ANTIGEN PROCESSING:
Cell engulfs an antigen bearing particle
LYSOSOME FUSES WITH ENDOCYTIC VESICLE
Endocytic vesicle forms
Particle is digested nto bit
MHC markers bind fragments of particle
ANTIGEN MHC complex becomes displayed on cell surface.
Cytotoxic T Cells will target intracellular pathogens, cells that have been infected INSIDE the cell and if it is a body cell then the Cytotoxic T Cell will release cytotoxins and destroy the cell.
MHC I - Are from inside the cell
MHC II - Outside the cell/body, external.
ADAPTIVE IMMUNE RESPONSE
Third line of defence are the lymphocytes
Specialised Lymphocytes
B Cells
T Cells
Depending on where the stem cell lymphocytes are matured, (T cells in thymus and B cells in bone marrow) they will become T/B cells
Pre-T cells leave the bone marrow and go into the lymphatic system/ circulation and then go to the lymph nodes, which is where they wait for antigens
Two types of adaptive immunity: Antibody mediated immunity/Humoural Immunity and Cell mediated Immunity
B cells → Humoral Immunity/Antibody Mediated Immunity → Plasma Cell which makes Antibodies, B CELLS do not make antibodies only the plasma cells.
T Cells → Cell Mediated Immunity → matures in the Thymus glance
Cytotoxic cells/CD8+ - will release cytotoxins
Helper T cells/CD4+ - will release cytokines
T Memory - A memory cell that will become either cytotoxic or helper T cells, for future infections
Regulatory Cell - T Supressor, stops the fighting
Natural Killer Cells - Lymphocytes - Not part of the adaptive immunity response, part of the innate immunity
B Memory - A memory cell that will become a plasma cell again
Cytotoxic cells have the CD8 glycoprotein and are activated by APC’s (antigen presenting cells)
They defend against intracellular bacteria, viruses, cancerous cells, transplanted foreign tissue, protozoa, fungi and worms.
Activated by MHC Class 1, releases cytotoxins such as perforin, granzymes to induce apoptosis.
Helper T Cells are not cytotoxic or phagocytic
They activate humoral immunity: B cells to make plasma cells to make specific antibodies
Cell mediated immunity: TC cells to release cytokines
Innate immunity: macrophages to initiate phagocytosis.
A naive Th-Cell means it hasn’t come in contact with that specific antigen before
Plasma b cells contain LOTS of rough er and undergo apoptosis
They make antibodies and antibodies are proteins so they need rough ER because ribosomes make proteins
Antibody Structure: Globular glycoproteins, quaternary structure
They are immunoglobulin
Two heavy and two light polypeptide chains (4 in total)
Chains are held together by disulphide bridges with Variable and Constant regions.
Order of amino acids determines the shape of the variable region binding site
Antibodies can neutralise antigens, agglutination (clumping bateria up), precipitation, assist in complement protein, enhance phagocytosis, inflammation, cell lysis
5 immunoglobulin isotypes
Humoural immunity
B cell receptor is the antibody
A naive B cell that has just processed the same antigen will present the antigen on its receptor
Lymphocytes
Circulatory system
Lymph nodes
Lymphatic vessels
Lymph fluid
Capillaries
The lymphatic system protects from infection and disease
Part of the immune system
Lymph fluid pass through lymph nodes
Network of lymph vessels connects the lymph nodes together
Lymphatic system acts as a one way drainage system, transports fluid from body tissue, houses lymphocytes and filters cellular waste
Capillaries are necessary so that all cells have access to oxygen and to get rid of CO2
Lymph Nodes, small structures that work as filters for harmful substances, located in strategic points
Immunity
Symptoms
Innate Immunity
Humoral Immunity
Cellular Immunity
Immunisation - harmless part of microbes are introduced to trigger the body’s immune response
Herd immunity
Vaccine - A suspension of antigens that are deliberately introduced into the body
Immunity can be
Innate or Adaptive
Natural or Artificial
Active or Passive
Natural Immunity occurs through contact with a disease causing agent, it is not deliberate; by chance
Artificially acquired immunity develops only through deliberate actions such as immunisation.
Both natural and artificial immunisation have the same result of activating adaptive immune response
Passive immunity is acquired through the transfer of antibodies, their immune system has not been activated, they are NOT receiving antigens, but instead antibodies. Memory cells are not made, they only have antibodies. Immunity only lasts as long as the antibodies are present.
Active immunity is the adaptive immune system activated. Delivery of antigens.
ACTIVE-NATURAL IMMUNITY
Pathogen/Antigen enters body (illness)
Adaptive immune system is activated
Memory cells are made
Long Term Immunity
ACTIVE-ARTIFICIAL IMMUNITY
About immunisation (vaccines)
Herd immunity
Active, deliberate process
Antigens are introduced and memory cells are made
Long term immunity
Types of Vaccines: Subunit or Whole agent vaccines
Subunit - contains some part or product
Whole-agent - contains whole, non virulent microorganism which can be inactivated (killed) or attenuated (weakened)
Herd Immunity, the resistance to the spread of a contagious disease within a population that result if a sufficiently high proportion of individuals are immune to the disease especially through vaccination
Minimisation of an epidemic
PASSIVE ARTIFICIAL
Antibodies injected
Used when a very rapid immune response is needed like antivenom
Human antibodies are injected
ANtibodies come from blood donors who recently had vaccination
Only provides short term protection
No memory cells
PASSIVE NATURAL
Mother’s antibodies pass across the placenta to the foetus
Colostrum (the first breast milk) contains lots of IgA which remain on the surface of the baby’s gut wall and pass into blood.
Immunotherapy
Metastases
Benign
Malignant
Cancer
Cancer is a group of diseases involving abnormal cell growth, with the ability to spread throughout the body
Benign or Malignant tumours
Beninghn cannot spread by invasion or metastasis
Malignant can spread through the body via the bloodstream
Immunotherapy uses Vaccines and monoclonal antibodies (mAbs)
Tumour cells thrive because they are able to hide from the immune system by expressive defective class-1 MHC
Immunotherapy uses the host system
SPECIFIC:
Marks cancer cells so immune system can find and destroy
Triggers response
B&T lymphocytes are stimulated to target cancer cells
Contain: Peptides, antigens or whole proteins of cancer cells and adjuvants
Adjuvants: substances that enhance the effect of a vaccine or other treatments
No side effects
Classified: Preventive or Therapeutic and personalised
Preventative, harmless virus-like particles containing viral DNA trigger immune response to create antibodies and memory cells etc.
Therapeutic: For the treatment of someone experiencing cancer. The tumour is not yet recognised by the immune system so immune cells are trained to recognise antigens and injected into individuals so the adaptive response is activated to be able to recognise and attack the tumour.
Monoclonal Antibodies are made to treat diseases
mAbs target specific antigens found on diseased or cancerous cells
mAbs can act directly when binding to cancer specific antigens to ;
Induce immunological response: apoptosis
Highlight cancer cells to immune cells
Block growth signals
Deliver toxins
NON SPECIFIC:
Boot immune system to work better
Trigger innate response, cytokines et: interleukin and interferon
AUTOIMMUNE DISEASES:
Diseases that have the immune cells targeting self-cells
Cytotoxic T cells, B cells activating, mast cells→ histamines and inflammation
No cure
Immune suppression that can help control overactive immune response and decreasing pain
Anti-inflammatory
Alongside immune suppressive medication
Acts as competitive inhibitor signals.
Interleukin competitive inhibitor, stops signals for immune system to work
Can stop the b cells, and t cells etc. Targets the immune system that will be targeting self-cells
Bacterial resistance
Antigenic shift
Pathogens
Population
Evolution
Reservoir
Novel Strain
Rational Drug Design - Targeted approach to designing new drugs, involves analysing the structure of a pathogen
Gram-positive - thick cell wall of peptidoglycan
Gram-negative - thin cell wall of peptidoglycan
Broad spectrum antibiotics - targets wide range of bacterial species
Narrow spectrum antibiotics - targets one or two bacterial species
Bacteriostatic
Bactericidal
Microevolution
Antigenic Drift
Antibiotics
Bottleneck Effect
Predation
Evolution = Change
Outbreak: the occurrence of one or several cases of a disease in an area in which it is not normally present
Epidemic: an uncontrolled outbreak that is the infection of many people simultaneously
Pandemic: An epidemic on a global scale, disease spread worldwide
Endemic: A disease that exists permanently in a particular region/population
Outbreaks require identification of cause, treatment, prevention of spreading and another outbreak
Condition for a pandemic
New pathogen/Novel strain - there is a lack of interaction with antigens and cannot be protected against because there is no immunity, no vaccine
Pathogen infects people and non-human hosts providing a ‘reservoir’
Pathogen is easily transmitted through direct contact, air or vector
Infected individuals are not isolated
No vaccination or preventative measures are in place
No control measures (masks, quarantine)
Influenza virus Structure
Two spike proteins, neuraminidase/sialidase and hemagglutinin
Nucleoprotein - RNA
Neuraminidase - enzyme, viral exit - cuts out
Hemagglutinin - viral entry, receptor - goes in
The virus enters and undergoes endocytosis, the nucleus is usesd for mRNA synthesis and RNA replication so now the virus has taken over the cell and will be used to make the virus while the virus exits and infects more cells
Subtypes of Influenza A are differentiated on the basis of the two surface antigens
Three Subtypes of H (Hemagglutinin) (H1,H2,H3)
Two of N (N1 and N2) generally cause the annual epidemics
Influenza A viruses are classified by A,B,C
Influenza A can cross species
A&B are main causes of epidemics/pandemics Type C cause mild versions
ANTIGENIC DRIFITING
There can be gradual minor changes in HA/NA caused by point mutation
Occurs in A&B
Vaccines are annually updated
ANTIGENIC SHIFT
Sudden Major Change, genetic reassortment of genes caused, when two subtypes infect a host, direct transmission from other animal to human introduces a NEW novel strain
Explosive Spread
Only in Influenza A
Creates a pandemic/epidemic
What are the scientific and social challenges presented in terms of the treatment strategies and vaccine programs (caused by viral antigenic drift and shift)?
Scientific: Vaccines created in response to viruses would have to change dramatically to accommodate for major changes due to antigenic shifts in the virus’s structure. Identification of mutation would be difficult and the difficult within identifying antigenic shift or drift.
Social: Immune responses of the population are naive and there is no immunity to a novel strain as the antigenic shift has drastically changed the virus’s structure. Vaccination development can be expensive and time-consuming as well as how measures will be implemented such as quarantines, lockdowns, masks, vaccinations etc.
CONTROL:
Antiviral drugs that prevent viral entry by binding to receptors
Inhibition of enzymes that catalyse reproduction of virus genome
Blocking transcription and translation
Prevents viruses form leaving cells to prevent further infection
RATIONAL DRUG DESIGN:
Rational Drug Design - Targeted approach to designing new drugs, involves analysing the structure of a pathogen
Uses this information to design a drug that will mimic or block the action of the disease-causing agent.
Produces drugs that have complementary shapes to the active sites of the pathogen or molecule they are targeting.
OTHER CHEMICALS TO CONTROL PATHOGENS:
Disinfectants - Non specific,
Antiseptics - Non specific, against bacteria, viruses and fungi
Antibiotics
Antibiotics:
Substances produced by microorganism or artificially that in low concentrations inhibits the growth or kills microorganisms
Broad spectrum antibiotics - targets wide range of bacterial species
Negative: Unnecessary introduction of antibiotics that may cause harm
Narrow spectrum antibiotics - targets one or two bacterial species
Better but takes time
Bacteriostatic - slows growth of bacteria by interfering with synthesis processes, like DNA replication, enzyme activity or protein synthesis
Bactericidal - kills bacteria, as an example may prevent the growth of cell walls to kill the bacteria.
Antibiotics only for bacteria because viruses hide in body cells and also they don’t have cell walls so bacteriostatic can’t target them, bactericidal can’t destroy the cell wall; they have different structures to bacteria
Four types of antibiotic resistance
Impermeable barrier, target modification, antibiotic modification, efflux pump mechanism
Evolution = Change in allele frequencies
Bacteria transfer genes easily and go into plasmids/DNA, high reproduction rate, exponential growth of population
Population: A population is the number of all the organisms of the same group or species, which live in a particular geographical area, and have the capability of interbreeding to produce fertile offspring.
Species: A group of living organisms consisting of similar individuals capable of exchanging genes or interbreeding.
Alleles: Alternate forms of a gene, two alleles is 1 genotype
Allele pool/Gene pool: The sum total of allele for all genes present ina population at one time
Large gene pool indicates genetic diversity and biological fitness
Small gene pool indicates low genetic diversity and biological fitness → Increasing chances of extinction
Used to determine allele frequency proportion of a particular allele within a population
Biological Fitness: The level of fitness a species has for surviving changes in environment based on the ability to adapt to new situations
Evolution change in allele frequencies over time
Mutation
Random change in the genetic composition due to changes in the DNA base sequence or chromosome
Point mutation
Gene Flow
Movement of alleles into or out of populations due to immigration/emigration
Gene flow keeps separate populations similar
Sexual Reproduction
Sex can introduce new gene combinations
Alter allele frequencies if mating is assortative
Random mating
Non-random mating
Assortative Mating - Preference for similar genotypes/phenotypes
Disassortative Mating- Preference for different genotypes/phenotypes
Genetic Drift
Completely random/chance
Causes the allele frequencies to drift from one generation to the next
No selective agents
May cause gene variants to completely disappear
Genetic drift has a greater effect on small populations (5/100 vs 5/10)
Population bottlenecks and Founder Effects
Bottleneck → Catastrophic event reduces population and reduces genetic diversity by chance
Founder Effect → Small group moves and reproduces in a new location, genetic variation is low
Gene Flow | Genetic Drift | |
Occurrence | Occur thru migration from one to another pop | Occur through random events. |
Population size | Larger population | Smaller population |
Reason | Inbreeding or inbreeding through migration | Sudden change or sampling error |
Evolution | Through migration | Thru bottleneck/founder |
Natural Selection/Selection Pressure
Change in gene pool composition as a result of differentially selective environmental pressures
Predation, Abiotic factors, Nutrition, Disasters, Finding a mate
Selective pressures are biotic and abiotic factors that select for certain characteristics in a population to be passed on and selected against other characteristics that will not be passed on
Eg. Dark bug camouflaging on a dark tree doesn’t get eaten by bird, bright coloured bug gets eaten
1. Variation: Intraspecies differences
2. Selection Pressure: A struggle is applied to population
3. Adaptations: Survival of the fittest
4. Reproduction: Adaptive quality is passed onto offspring
5. Change in population: Allele frequency/microevolution
Increase offspring of a certain trait due to survival of the fittest (organisms most capable of reproducing produce more offspring with whatever trait they have that has enabled them to survive and reproduce)
If there is no selective pressure, different traits do not matter
There needs to be a struggle to live (selective pressure)
Selective pressures:
Just because a phenotype has been wiped out does not mean the allele has disappeared/genotype may still have the alleles that will trigger the recessive allele
The allele cannot be dominant because then even a single allele will trigger the phenotype thus, if the allele IS dominant it has gone extinct
NO SELECTIVE PRESSURE NO CHANGE!
Things that mean NO change to allele frequencies: Large population, no mutation, no migration, random mating (organisms choose partners randomly), no selection (no traits can be favoured)
Point mutation
Block mutation
Chromosome
Meiosis
Allele
Gene sequence
Mutation
Substitution
Deletion
Addition
Non-disjunction
Adds a new allele to increase gene pool
A mutation is a change in the gene sequence or chromosome.
Gene mutations (point mutation) localised changes to DNA base sequence ie: substitution, deletion or addition
Chromosomal Mutations (Block mutation) large scale mutations occurring during meiosis that can change chromosome structure and number
Gene mutation:
Missense substitution
Type of mutation is a change in one DNA base pair resulting in amino acid subbing in for another in the protein made by a gene
Eg. Sickle Cell Anaemia
Nonsense Substitution
Accidental mutation codes for STOP codon resulting in a shortened protein/junk protein
Insertion (Frame Shift)
An insertion changes number of DNA bases, protein may not function properly as point mutation has caused a frame shift
Silent mutation:
Nothing is changed, redundancy allows for the same amino acid to be produced
Deletion (Frame Shift)
Changes number of DNA bases, frame shift because protein will be changed sequence
Frameshift Mutation
Can change every amino acid that follows point of mutation and alters a protein so much
Changes the sequence, shifts reading frame
Chromosomal Mutations [Block Mutation]
Block mutations can cause polyploidy, changes in chromosome number and structure.
Duplication
Alleles are duplicated/added
Inversion
Positions are swapped, frequency doesn’t change but sequence does
Deletion
Removed segments of chromosome
Insertion
New alleles introduced
Translocation
Also swaps but with a different chromosome
Meiosis:
Meiosis can stuff up and nondisjunction may occur, Main cause for aneuploidy
Aneuploidy, incorrect chromosome number
(Chromosomes do not separate properly)
Variation is introduced because random alignment and crossing over/random assortment when there is exchanging of whatever
Polyploidy cells are organisms containing three or more times the haploid number of chromosomes like 3N or 4N, uncommon in animals, common in plants
Occurs through allopolyploidy and autopolyploidy
Allopolyploidy - An individual or strain whose chromosomes are composed from two different species to produce hybrids
Can have the full chromosome set of two different species
Autopolyploidy - resulting in offspring with two sets of chromosomes from it’s own species
Criteria: variation in a population and heritable traits
Humans select
Determine desired trait
Interbreed parents with desired trait
Select offspring with desired trait and interbreed them
Process continues until reliable reproduction of desired trait is achieved
Problems with selective breeding
Gene pool has been reduced and alleles are lost therefore reduced resistance to environmental change
Reduced Biodiversity Genetic diversity decreases, ability for adaptation is decreased
Increased genetic abnormalities, Genetic defects can be selected for with favourable traits
Stratigraphy
Geologic Time Scale
Palaeontology - the study of fossils
Body fossil
Trace fossil
Impression fossil
Mineralised fossil
Intermediate
Transitional fossil
Permineralisation
Index fossil
Sedimentary
Strata
Geologic Time Scale (GTS) is a system of chronological dating that relates geo strata (stratigraphy) to time
First prokaryotic life form - Cyanobacteria
Fossils preserved in rock, soil or amber
Remains of organisms
Palaeontology is the study of fossils
Lowest rock layers are older
Rock layers formed later contain more complex organisms
Variety also increases
Body Fossils - fossilised remains of an organism eg: bones, leaves
Trace Fossils - No parts of an organisms, impressions of activity eg: footprints
Impression Fossils - Organism decays and leaves an impression the rock/earth
Mineralised fossil - minerals replace the organism structure
Fossilization requires
Rapid burial: protection against scavengers, erosion and damage
Low oxygen: protection of oxygen damage and lack of decomposition
High pressure: to promote mineralisation of remains
Remains undisturbed: To allow for permineralization
Hard body parts: eg. teeth, shell
Death and decay: Soft body parts decay, leaving only hard body remains
Deposition/rapid burial - hard remains are rapidly covered with silt and sand and layers build over time
Permineralization - pressure from layers od dirt and rock cause hard organic material to be replaced by minerals
Erosion/ exposure - movement of earth plates may displace the fossil and return to discovery
Soft body fossils are less likely because they have more water and are more likely to decompose, more likely to create impression fossils (ie: jellyfish)
Transitional fossils: Fossil that can link two different lifeforms, remains of a pre-existing organism that shows a progression/transitions
Should show transitional/intermediate characteristics
REMINDER: GO THROUGH PRESENTATION 6. FOSSILS WE LOST TIME AND GO THRU IT BETTER
Absolute dating: anything that gives exact dates and numbers, numerical dating, determined by radiometric dating, estimates the age in years by measuring certain radioactive isotopes the object contains.
Relative dating: estimates the age of fossils found within strata, cannot tell the actual age of the fossil, using index fossil/stratigraphy, older than this/that, MUST COMPARE SOMETHING
Sedimentary rocks
Some of these layers may be laid down by water (Sedimentary) or volcanic activity (igneous)
Importance of the sequence in which is was deposited etc.
Relative dating is important to figure out index fossils
Index fossils are organisms that were geographically widespread and abundant but only existed for a limited span of time.
Must be distinctive, globally widespread and recognizable, became extinct quickly to pinpoint precise time periods
ABSOLUTE DATING
Radiometric:
Using the isotopes of carbon to determine age
Isotopes: Variation in neutrons
The presence of Carbon-13 and Carbon-14 indicate how long something has been around and the decay of that isotope
Isotopes go through radioactive decay
Parent Isotope is unstable
Daughter Isotope is stable
Radioactive decay is when the parent isotope becomes stable
When half of parent isotopes have decayed/become stable becomes 1 half life and so forth
Carbon-14 is not old enough for REALLY OLD shit
Carbon 14 takes 5730 years for half of the isotopes to become stable and become N14
Process of C-14 → N-14
Cosmic radiation heat N-14
N-14 loses a proton → C-14
C-14 + C-12 are in the atmosphere and get absorbed by living organisms
When those organisms die, bones lose C-14 as it becomes N-14 via beta decay (gains a proton)
Maximum limit of this method is 60 000 years
LIMITATIONS OF FOSSIL RECORDS
Organisms decompose rapidly
Are eaten
Soft-bodies organisms do not fossilise easily due to water
Small fraction of organisms die in conditions favourable to fossilisation
Fossils are still unearthed
Macroevolution
Speciation - the evolution by which new biological species arise over time
Species - Organisms that can produce fertile offspring with one another excluding asexual reproducing organisms
Ancestral population
Allopatric Speciation
Prezygotic Barrier
Postzygotic barrier
Geographical isolating mechanism
Isolating mechanism
Reproductive mechanism
Adaptive radiation - divergence of a large number of related species from a common ancestor
Speciation is MACROevolution - the evolutionary process by which new biological species arise over time
Ancestral populations are divided then isolated preventing gene flow
Different selective pressures will create differences in population
Allopatric speciation
Ancestral population: There is gene flow and variations
Isolating Mechanism: prevents gene flow (the movement of genes between populations)
Geographical isolation
Reproductive/Genetic isolating
Mutation creates new variants in different areas
Natural Selection: different selection [pressures select for new vairants
Speciation: individuals from each population can no longer produce fertile offspring with each other or the original ancestral population.
Reproductive Isolation
Mechanisms that prevent mating and reproduction, ie: individuals not responding to courtship
No gene flow
No exchange of alleles between populations
Prezygotic isolation
No fertilisation → No zygote
Postzygotic isolation
Zygote is formed but it is inviable or infertile (dies or can not reproduce)
Pre-Zygotic
Post Zygotic:
Adaptive radiation - divergence of a large number of related species from a common ancestor
CHARLES DARWIN'S GALAPAGOS ISLAND FINCHES:
Finches with different features found on Galapagos islands all descendants of the mainland ancestral species.
Geological Isolation
Sympatric Speciation
Species share the SAME geographical area but are reproductively isolated
Isolation comes from within the group
Assortative mating
Relatedness
Relation
Divergence - Speciation
Homologous Structures
Adaptive radiation
Vestigiality
Molecular homology - sameness on a molecular level
Comparing anatomical structure, Mitochondrial DNA, genetic sequences
Structural vs Molecular
Homologous Structures
Evolved from the same structure in an ancestral species
Different functions
Immediate common ancestors
Organisms show divergent evolution
Adaptive radiation
Eg. Pentadactyl limb
Vestigial - Describes homologous characters of organisms which have seemingly lost all or most of their original function in a species
Vestigiality can be structures, behaviours and biochemical pathways
Changes to the environment have rendered these structures redundant and so over time they have lost their functionality
Same function
Different structure
No immediate common ancestor
Organisms develop the same structure due to convergent evolution
Molecular clock hypothesis is:
Changes in DNA and proteins are constant over evolutionary time and across different lineages
The amount of molecular change between two species measures how long ago they shared a common ancestor
Molecular clock calculations are carried out on DNA or amino acid sequences btw species to establish relatedness
Comparison of DNA sequences and amino acid sequences
If divergence has occurred further back in time, there will be less similarity/less molecular homology
Mutation Rate:
Change in DNA over time
Can be expressed as the number of nucleotide changes over a million years
Molecular clock uses the rate of accumulation of mutations of DNA to determine how long ago divergence occurred.
Less Related Species:
More mutations
Greater differences
Divergence occurred further back in time
Closely related species
Less mutations
Less differences
Divergence occurred more ‘recently’
Amino acid:
Differences in amino acid sequence reflect changes in DNA sequence
Changes in the gene nucleotide should build over time
DNA comparison:
Direct Comparison of DNA base sequences
Comparing whole genome
DNA hybridisation
Comparing karyotype
Mitochondrial DNA
Mitochondrial DNA
Only inherited from mother
Mostly for recent (20 mil)
Can be recovered from teeth and bones
There’s a lack of recombination of mtDNA, remains the same
Higher mutation rate: Contains non-coding regions known as the D-loop that mutates at a higher rate
High copy number - cells have lots of mitochondria
Maternal inheritance: mtDNA is inherited from the mother only, to establish ancestry • It is used as a molecular clock.
Mitochondrial Molecular Clock: rate at which mutations have been accumulating in the mitochondrial genome of hominids during the course of human evolution.
Closely related species will show more similarities in base sequences, genome, DNA, Chromosomes, mtDNA
Lineage
Phylogenetic
Ancestor
Ancestral lineage
Descendant
Relatedness - determined by comparison of aa sequences or DNA to establish phylogeny, how recently species diverged
Phylogeny - refers to an evolutionary line of descent, can be determined by comparing sequences in different species.
Phylogenetic trees act as evidence for relatedness
Phylogeny refers to the evolutionary line of descent, determined by comparison
Phylogenetic tree:
Hypothesis relatedness
Phylogram
Compares sequences that have a constant rate of mutation (evolutionary clocks)
Mitochondrial DNA is a useful source as it is maternally derived has a known mutation rate and lacks recombination
Difference → mutation in either nucleotide of amino acid sequences.
Evolutionary trees can evolve to alter hypotheses if new evidence alters understanding
All living organisms have Cytochrome B
Hominidae
Genus
Family
Order
Species
Hominoid
Primate
Primary ancestors would have had
Arboreal
Grasping hands
Long, mobile limbs
Quadrupedal locomotion
Binocular vision
Upright sitting position
Nails instead of claws
Large eyes to improve eyesight, colour vision
Large highly developed area associated for vision
Reduced development for smell
Different types of teeth for wider variety of food sources
Singular birth: Longer parental care, increased infant dependency
Tails
Hominoidea - superfamily that includes apes and humans
Hominoids - members of the superfamily hominoidae
Hominids - all modern and extinct great apes. Gorillas chimps etc. and Immedieate ancestors
Hominins - any species of early human that is more closely related to humans that chimpanzees including modern humans
Pre-Hominins - Arboreal lifestyle, food resources were readily available in near-continuous forest
Cooling climate —> Trees became scarce
As trees became scarce, pre-hominins were forced to leave trees in order to seek out food sources
Bipedal Walking
S-curved spine
Inward femur angle
Pelvis shape
Foot shape/structure
Reduced Canines (Not related to walking)
Foramen Magnum - hole in skull
Brain size/Skull
Bipedal motion through S-shaped, flexible spine for balance
Femur/tibia angled inwardly for centre of gravity and allow for balance and walking
Short broad pelvis to allow for attachment of large powerful muscle
Bow shaped to support torso organs
Chimpanzee feet has opposable thumbs
Human feet has an arch, acts a spring, loss of big thumb, slightly larger toe
Transverse Arch - Converts foot into a spring allowing for transmission of stresses and improving walking efficiency ef
Foramen Magnum is the hole at the base of the skull thru which the spinal cord passes
If the foramen magnum is positioned towards the back (posterior) - quadruped
Foramen is more centred/to the front – Bipedal
Larger brain size, reduced brow ridge and flatter face
Cranium Capacity - Mass of brain that can fit into a human
Trends in Skull Anatomy
Shape and slope of forehead
Brow ridge
Facial Angle
Size of teeth
Protrusion of mouth
Position of foramen magnum
Size and shape of zygomatic arch
Brain case - size and shape
Size of mandible
Sagittal crest present? (Humans, not present)
Shape of occipital region
Differences between skulls of Australopithecus Afarensis and Homo Sapiens
Differences | Austra | Homo Sapiens |
Size of Skull | Smaller skull
| Larger skull |
Brow Ridge | Has one | Doesn’t have one, not very prominent |
Foramen Magnum | In the posterior, towards the back to allow for quadrupedal posture | In the anterior, centre of skull so head is atop spine and balanced |
Teeth Size | Larger teeth
| Smaller teeth |
Jawbone | Wider Jawbone | Narrower Jawbone |
Face Slope | Slanted slope, on an angle, teeth are jutted out and jaw recedes at an angle | Protruding jaw but straight face slope/vertical |
Zygomatic Arches | Very prominent cheek bones | Reduced |
Arms | Longer than legs, used for walking | Shorter than legs, not used for walking |
Why was Bipedalism Selected for/Adapatations!
Enabling a more proficient use of tools by freeing hands when upright
Bipedalism is more energy efficient
Thermoregulation: Lowers body temperature as solar radiation is retatined
Greater view of surroundings therefore able to escape predators more effectievly
Greater ability to disperse and cover more ground leading to habitat variability
More effective mating strategies leading to successful reproduction
Reduced canines was also selected for
Ability to carry food and weapons when walking
Carrying offspring while moving and eating
Hair loss
Retention of head hair for reflection of heat
Easier to control parasites
Thermoregulation - Less trapped heat, Greater heat loss, well developed sweat glands
Early Hominins
Australopithecus
Homo genus
Environment, what they looked like, how they moved etc.
Homo Erectus
First human emigrant
Left Africa
Founder effect, some remained in Africa
Larger brains and advanced toolmaking
Left btw 100 000 and 1.6 million years ago
Wore skins enabling travel from Africa into china and south East Asia
Homo Floresiensis
‘Hobbit’
Small stature
Wide pelvis and hunched shoulders
Lived in Asia between 100,000-60,000
Flat face’
Theories
Smaller body possibly due to surviving with constrained resources, island dwarfism
Descendants of homo habilisi
Has a pathological condition in modern humans (microcephaly)
Homo Neanderthalensis
Complex species
Bigger brain than humans
Similar but they have a bigger brow ridge
Shorter
Had a more robust skeleton and muscular
Cousins NOT direct ancestors, shared a common ancestor
Co-existed with modern humans
Went extinct 28000 years ago
Shows many single base differences however DNA is very very similar
Neanderthals did not contribute any mitochondrial DNA to any homosapiens living today
Human females and Neanderthal males
0% of Neanderthal DNA in African populations
1-4% in people of European or Asian descent
Did not migrate to Africa, only came in contact in Eurasia
Possibly went extinct due to transmission of disease, pressure of incoming migrant humans, or being slaughtered by modern humans (killer ape theory)
Homo Denisovans
Diverged from modern humans about 500 000 y.a., related to neanderthals
Diverged from neanderthals about 300 000 years ago
Possibly resembled neanderthals
Epigenetics:
adding methyl groups, not actually altering DNA etc
Working out pattern of methl or whatever could see gene expression and effects on appearance.
Archaic Human
Includes neanderthals, H. Floresiensis and H. Denisova
Mainly differed in skull
Backward sloping forehead
Big brow ridge
Long elongated skull
Why did they leave Africa:
Depletion of resources
Competition for resources
Climate change - droughts leading to starvation
Curiosity
Human Migration Evidence
DNA Evidence to support the hypothesis that early hominins migrated out of Africa around 150 000 ya
DNA evidence suggests interbreeding btw modern homninins and nanderthals in Europe and the Middle East
Reached australia 35-60 000 ya
The Pleistocene Ice Age created a land bridge that connected Asia and Alaska over 13,000 years ago.
DNA suggests that modern humans reached australia continent approx 55 000 ya
Out of Africa = Replacement theory
H. Erectus left, evolved into Archaic humans
H. Sapiens left 200 00 years later and replaced archaic humans
Out of Africa model II = Assimilation theory
Same as A except some H. erectus survived and interbred and joined H. Sapiens
Multiregional - Continuity theory
• Significant migration of H.erectus across Africa, Asia and Europe for the last 1.8my. • Isolation of the populations à divergence of gene pools, traits & behaviour. • interbreeding btw populations could have occurred • genetic drift led to the DNA of the other species being lost from the H.sapien genome.
Multiregional theory 2
Haplogroup
Group who share common ancestor on paternal or maternal line
Inherited Y chromosome
Inherited mtDNA
Indigenous Migration
Single migration from Africa to Australia 60 000
42 000 ya, extinction of megafauna
Aboriginal people have strong sense of country and place
Place- space mapped by intangible boundaries that individuals or groups of Torrest Strait Islander peoples occupy and regard as their own via spiritual and emotional connections
Connections:
Connection via Maternal and paternal lines of descent • Clan group • Language groups • Spiritual connection