dna and genetics + biotech

plasmid

small circles of dna, a genetic structure in a cell that can replicate independently of the chromosomes

restriction enzymes

enzymes that cut DNA and specific restriction sites found naturally in bacteria

sticky end

the same enzyme used on e coli is the same one used to cut DNA so plasmid and DNA have the same

e coli cell wall

negatively charged and repels negatively charged molecules

positive calcium

neutralizes the negative charge on cells outer membranes, allows DNA to cross the plasma membrane

heat shock

a sudden increase in temp causing outside pressure to increase, allows plasmid DNA to enter the cell

bacterial conjugation

a plasmid is shared between 2 bacterial cells

transformation

bacteria take up DNA, including plasmids directly from the environment

ori site

where DNA polymerase initiates replication

Luria broth

a medium that supports bacterial growth

selectable markers

only bacteria having the gene will survive in presence of antibiotic

culture

isolated population of cells

colony

a clump of bacterial cells that have grown and divided

p+

competent cells with plasma

p

- competent cells with no plasmid

label two new tubes, P

and P+

plate 50 uL of P

and P+

LB

nutrients

LB/Amp/Ara

nutrients, antibiotics, arabinose

15 minutes

the amount of time tubes are kept on ice after adding plasmid to tubes

45 seconds

the amount of time tubes are placed in hot water

1 minute

the amount of time cells recover after coming out of hot water

150 uL

the amount of LB broth added to each tube after recovery on ice

50 uL

the number of microliters of cells added from tube to plate

PCR

used to quickly identify a specific DNA sequence and then make multiple copies

DNA

becomes single-stranded at high temperatures because its two strands separate and denature

primers

short single strands of DNA stick to specific sequences of DNA and can only occur with complementary bases

DNA polymerase

an enzyme used to replicate DNA

taq dna polymerase

can function at higher temperatures

denaturation phase

DNA sample containing the sequences to be amplified and splits at high temperatures

annealing phase

single-stranded DNA primers annealed to denatured single-stranded DNA at a cooler temperature

extension phase

taq polymerase replicates the region of interest by adding nucleotides to the 3 end of the primers

thermocycler

controls temp and length of time for reaction

gel electrophoresis

a technique used to separate DNA fragments according to their size and charge

indentations

wells where DNA samples are loaded and pulled through an electric current to the other end of the gel

DNA fragments

negatively charged moving towards the positive electrode, and have the same amount of mass charge

bands

A well-defined "line" of DNA on a gel

yardstick

DNA fragments of known sizes that help determine the absolute size of an unknown DNA fragment

agarose

a polysaccharide that makes gels to separate DNA, and comes in dry padded flakes

wells

pocket-like indentations where DNA samples are placed

gel box

where the gel is placed before DNA samples are placed

buffer

can conduct the current of the gel

DNA ladder

contains known DNA length

lane

corridor through which DNA passes as it leaves a well

genetic engineering

a branch of biotechnology that uses special procedures and techniques to change an organisms DNA

rfp gene

the red fluorescent protein in sea anemones

proto-oncogenes

a gene that tells the cell to move forward in the cell cycle, will continue the cell cycle if mutauted

tumor suppressor genes

slows down the cell cycle and tells a cell when to die, will cause the cell to live longer if mutated

P53

cell protein that commonly mutates to allow cancer, allows cells not to die when mutated

benign tumors

slow growing, capsulated, noninvasive, do not metastasize, well-differentiated

malignant tumors

fast-growing, nonencapsulated, invasive and infiltrate metastasize, poorly differentiated

cancer

a disease caused by uncontrolled growth of cells

cell cycle

the process of cell growth and division

interphase

period of growth that occurs between cell divisions

cell division

the stage in which the cell divides, and the cell grows, prepares for division, and divides to form two daughter cells

G1

cell grows in size and organelles are duplicated

S phase

DNA replication occurs

G2

additional cell growth

M phase

cell divides into two daughter cells

dna ligase

reseals dna strands back together

mitosis

division of the nucleus within a cell

cytokinesis

division of the cell cytoplasm

prophase

spindles are formed, chromatins condenses into chromosomes

spindle

fan like structure that separates chromosomes

metaphase

chromosomes line up across the center of the cell, and microtubules from the spindle connect to each chromosome

anaphase

chromosomes are split in half

telophase

chromosomes gather at opposite ends of a cell, and the nucleus splits

cytokinesis

cytoplasm pinches in half to make 2 separate cells, and each daughter cell has an identical set of chromosomes

checkpoint control system

indicates whether key cellular processes have been completed correctly

G1 checkpoint

determines whether dna replication can begin

G2 checkpoint

checks to see if dna synthesis had been done correctly, signals that mitosis can begin

M checkpoint

checks spindle to make sure that they are all attached to a chromosome and makes sure that they can separate properly

replication

cell makes a copy of its DNA before it divides, occurs during the s phase and ensures that each resulting cell will have a complete set of DNA

helicase

an enzyme that unzips a molecule of DNA

primer for DNA

a short sequence of DNA that allows for DNA replication to begin

messenger rna

carries dna message from nucleus to cytoplasm and tells ribosomes which proteins to make

ribosomal rna

makes up ribosomes which are organelle that assmebles amino acids into proteins

transfer rna

attaches to mrna from a dna template creating a chain of amino acids that will form a protien

transcription

process of making mrna from a dna template, a single strand of mrna is formed by reading dna

translation

process of converting mRNA into a sequence of amino acids that build into a protein and happens in the ribosomes

codon

3 mrna nucleotides code for an amino acid

nucleus

where the dna is located in the cell

DNA

deoxyribonucleic acid, holds all of our cells information, instructions to make all the proteins in our bodies

T

thymine

A

adenine

C

cytosine

G

guanine

body is made up of trillions of cells

each cell is a copy of a single cell that divided itself to make all of the cells in your body

cells

need instructions to create who you are

gene

specific part of dna that gives body instructions for making protein

chromosomes

structure in the nucleus that is made up tightly wound dna, humans have 23 pairs of chromosomes

double helix

structure of dna

multipotent

can change into other kinds of cells but not all

stem cells

a specific type of cells capable of evolving into many types of specialized cells within the body

embryonic stem cells

supply new cells for an embryo as it grows and develops into a baby

adult stem cells

supply new cells as organism grows and replace cells that get damaged

chromatid

half of a chromosome

chromatin

strand of dna and proteins

centromere

where the chromosomes connect to the microtubules

pluripotent

can evolve into any cell type