Cloning and Biotechnology

what is a clone

offspring produced by mitosis that is genetically identical to its parent plant

what does asexual reproducting mean

the generation of new individuals, using mitosis to produce clones , naturally

what is reproductive cloning

using artificial cloning methods to produce 2 or more individuals that are clones of each other

what does vegetative propigation mean

the production of plant clones from non reproductive tissue.

what does perennating organ mean

plant structures which allows them to survive in adverse conditions. Contain stored food and can remain dormant in soil

what does horticulture mean

branch of agriculture that deals with plants

what does agriculture mean

the breeding if animals, plants or fungi for food or other resources

what does taking cuttings involve

removing and planting short sections of stem of a plant in order to produce clones

give examples of natural ways of vegetation proprigation in plants

bulbs, runners, rhizomes, stem tubers

why is natural cloning in plants very common but not in animals

plants have many totipotent cells but animals do not

give artificual ways of vegitation propigation

cutting, bulb splitting, rhizome splitting

which crops are commonly propagated by cloning

banana, sugar cane, sweet potato

what ways can you improve vegetative propigation by cutting

• use a non flowering stem

• use hormone rooting powder

• keep cuttings well watered

• create a humid enviroment

what does tissue culture mean

the method of growing plant cells, in isolation from the parent plant under sterile conditions in or on a nutrient culture medium of known composition

what does micropropagation mean

the process of making large numbers of genetically identical offspring from a single parent plant using tissue culture technique

what does explant mean

the material removed from a parent plant for tissue culture

what does callus mean

a mass of undifferencated plant cells that have been grown from an explant

outline the process of micropropagation by callus tissue culture

• take meristem of the plant you want to clone

• sterrilsed using chemicals, bleach or ethanol

• then is added to a petri dish containg a balance of plant hormones, which stimulate plant growth

• callus transfered to a new environment containg hormones and nutrients which stimulate development/ or into plantlets

• then planted

why is microptopagation used over sexual reproduction

• if plant does not readily produce seeds

• desirable plant is very rare

• can produce very large number quickly

why is micropropagation used over vegatative propagation

• plant might not respond well to natural cloning

• if its required to be pathogen free

what are some advantages and disadvanteges of tissue culture

• quick and can save almost extinct plants, all times of the year

• expensive, could transfer diseases, easier for crops to get a disease

what does binary fission mean

the method of aseual reproduction in bacteria

name the natural cloning methods

mitotic parthenogenesis

damage

monozygotic twinning

budding

fragmentation

what does SCNT stand for

somatic cell nuclear transfer

what does natural cloning mean

when animals produce genetically identicle offspring using asexual reprodcution

outline natrual cloning in invertebrates (worms)

• fragmentation/ regeneration forming new genetically indentical offspirng

• buds break off the parent organisms and grwos in to the new genetically identical offspring

outline natural cloning in verterbrates

• when an early embryo splits to two genetically identical embryos

• each grows independently, so genetically identical offspring so monozygotic twins formn

what is a monozygotic twin

genetically identical offspring that come from the same embryo that then splits

• they are identical twins

what is artificial twinning

process of splitting an early stage embryo into multiple embryos which are then implanted into a surrogate

what does artifical twinning involve

• parents with ideal characteristics are bred

• female given hormones to produce more mature ova

• used for IVF

outlie the process of artificial twinning

• fertilised egg undergoing early cell division to form an embryo

• embryo then divides into 2 half embyros

• embryos then deveopl further in a lab before being implanted intp mother

• then 2 identical twins are born

outline the process of SCNT

• somatic cell is taken from one animal to be cloned and the nucleus is removed to be cloned

• then an unfertilised egg from an egg donor is selected and the nucleus is removed

• they are then fused together by an electric shock which stimulates the devision

• its then added to a surrogate mothers uterus

na,e some uses of animal cloning

• medical research

• conservation

• ariculture

• pharming

• stem cells

what are some pros of animal cloning

• animals with good characterisitcs can have lots of offspring

• SCNT can reporduce endangered species

what are some cons of animal cloning

• high miscarriage rate during implantation or malformed offspirng

• animals produced have shorted life span

what does biotechnology mean

how we use living organisms or their products so that they are usfel to us

which are the most commonly used organisms in biotechnology

bacteria and yeast (fungi)

why are microrganimss used for biotechnology

• grow rapidly

• growth no dependent on season

• less pollution

• grow easily on inexpenisve materials

• production costs are low

• easier to carry out selective breeding

• few ethical pr welfare issues

• can be grown anywhere

• products made quickly

what are problems with using microgorganisms 

• can be infected by viruses resulting in production halting 

• human objections to eating food made out of waste materials 

• a lot effort required to prevent contamination from other microorganisms 

what are some advantages of using microrganisms in human food

• can be made to taste like anything

• have a low fat content

• have high protein content

• reproduce quickly so produce protein faster than animals or plants

• suitable for vegetarians

• produce protein with a similar amino acod profile to animal and plant protein

• have low cholesterol content

what are some negatives of using microorganisms

• have little natural flavour

• costly to seperate microorganisms from the material on which they grow

• can produce toxins if growing conditiond are not optimal

• microbial biomass can have a high proportion of nucleic acids 

• texture is different to traditional protein sources 

what does culture mean

growing microganiss in liquid nutirent broth or on a solid surface

pure culture meaning

single species (quorn)

mixed culture meaning

mixture of species (yogurt)

what features does a commercial fermenter have 

stainless steal vessel walls and whyv

dosent rust

water jacket ?

control the temperature easily for optimum growth

paddle stirrers

avoid clumping and ensures nutrients are available to all

sparger 

mix culture and oxygenate 

nutrient input

provided to ensure good growth

temperature probe

monitor temp for optimum growth

pH probe

optimum pH levels

harvest pipe 

release culture at the bottom because of gravity 

gas outlet

so that there isnt a build up of gas pressure

oxygen prope

optimum O2 level for anerobic or aerobic respiration

how is batch fermentation carried out

• microorganisms inoculate into a fixed volume of medium

• as the culture reaches the stationary overall growth ceases but during this phase the microorgansims often carry out biochemical changes to form the desired end products

• as growth takes place, nutrients are used up and new biomass and waste products build up

• the process is stopped before the death phase and the products are harvested. The fermented is cleaned and sterilied and new fermenter set up

• population goes through lag, log and stationary phases of growth

how does continous fermentation work 

• microorganisms are inoculated into sterile nutrient medium and start to grow 

• sterile nutirent medium is added refularly to the culture once it reaches the exponential point of growth 

• culture broth is constanly removed keeping the culture volume in the bioreactor constant 

• product harvested regularly throughout the process 

• maintain population in log phase of grwoth 

what two types of metabolites are there and what do they look like on a graph ?

primary and secondary

what features do primary metabolites have?

• produced in favourable growth conditions

• mostly produced in the lag and exponential phases of growth

• produced as an essential part of growth

• glucose and enzymes are examples

what features do secondary metabolites have 

• help microorganisms survive unfavourable conditions 

• produces in conditions that are less favourable for growth 

• not essential for growth 

• mostly produced in the stationaty or decline phases of growth 

• antibiotics 

extracellular

enzymes that are secreted from the microorgansims into the culture medium and catalyse reactions outside the cell

immobilised

enzymes that are bound chemically or physically to an inert substance

intracellular

enzymes that are containe witin the microorganism and catalys reactions inside the cell

isolated 

enzymes that are not contained within a cell 

what methods of immobilisng enzymes are there

chemical: absorption ionic bond, covalently bonded

physical: membrane seperation, lattice entrapment in gel bead

what advantages are there of using immobilised enzymes

• enzymes can be reused

• enzymes easily seperated from reactants so products arent contaminated

• more tolerant of temperature variations

• less likelt to be denatured by pH changes

• allows continius processing

what disadvantages are there of using immobilised enzymes

• enzymes active site may be altered by immobilisation

• higher intial costs of materials

• higher intial costs of bioreactor

• large diffusion barrier

• enzyme may be less free to collide with substrate

absorption method

• enzymes are mixed with an immobilisng support and binds with ionic links

• binding forces are variable

• active sites can be hard to access if ionic bond is facing the wrong way

• small diffuison barrier

covalently bonded method

• difficult to do, enzymes are covalently bonded to an insiluble support using cross linking agents

• strong binding force

• acitve site is easy to reach 

lattice entrapment method

• difficult to achieve, enzymes are trapped in a gel bead or network of cellulase fibres

• weak bonds, which causes leakage

• active site not easy to reach due to mesh surrounding it

membrane separation method

• simple to do, partial permeable membrane may separate enzymes, solute flows past reactants diffuse through

• there are no bonds, no leakage

• hard to reach active site, hard for sufficient substrates to diffuse through the membrane to reach the enzymes 

what si the liquid culture medium and solid culture medium

liquid= broth

solid= agar

ways to count growth of culture

• direct counting

• viable counting (one you did)

• turbidiemtry