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Biology Module 6 > 6.4 Cloning And Biotechnology > Flashcards

6.4 Cloning And Biotechnology Flashcards

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1
Q

What are clones

A

Genetically identical organisms or cells

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2
Q

How can clones be produced

A

asexually (mitosis, budding, binary fission) or sexually

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3
Q

Advantages of natural cloning

A

if the condition is good for the parents it will be good for the offspring = high yield of the product, cloning is fast, reproduction can happen even in the absence of parents = good if it’s a rare species

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4
Q

Disadvantages of natural cloning

A

through natural cloning areas can become overcrowded by offspring, no genetic diversity unless there’s mutations = little variation, if something happens that wipes one of them out it’ll wipe all of them out

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5
Q

What does plant cloning happen using

A

Their vegetative parts = aka vegetative propagation = natural cloning in plants involves this

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6
Q

Examples of how vegetative parts of plant are creating clones:

A

Runners/stolon, suckers
Bulbs, corms
Tubers

how natural cloning occurs in plants bc it gives rise to new plants that are genetically identical.

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7
Q

Runners/stolon

A

when the stems grow on the surface of the ground

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8
Q

Rhizome

A

If the stems grow underground

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9
Q

Sucker

A

extension of the roots underground

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10
Q

Tubers

A

underground stems e.g. potatoes = example of plant natural cloning

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11
Q

Natural cloning in animals overview

A

When zygote splits into 2 daughter cells + produces twins

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12
Q

Natural cloning in insects

A

not common but water fleas and green flies can reproduce asexually.

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13
Q

Plant cuttings

A
  • to get a plant cutting you have to cut the stem at two nodes (internode contains meristem tissues)
  • The stem should then be placed in moist soil and new roots will form
  • Some plants will naturally form roots, some may need help so you can add rooting powder (contains rooting hormones)
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14
Q

Where else can plant cuttings be taken from

A

Root, leaf

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15
Q

Larger scale version of plant cuttings

A

Tissue culture (vegatative propagation

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16
Q

Vegetative propagation

A

concept of growing cells, tissues or organs from a small sample of cells/tissues

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17
Q

To make tissue culture large scale

A

Micro propagation

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18
Q

Example of piece of plant tissue is used for Micropropagation

A

a leaf (best piece to use is meristem because it is free from viruses)

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19
Q

Callus

A

small lump of undifferentiated cells

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20
Q

Outline Micropropagation

A

-take a small piece of plant tissue
- Cut this into smaller pieces + call these pieces the explants
- Take the explants and sterilise them using bleach or alcohol
- Place the explant on a sterile growth medium e.g. agar containing suitable nutrients including glucose, amino acids, auxins and cytokinins = stimulates mitosis in the plant so you’ll see some growth = initial growth is known as a callus
- Take the callus and move it to a new agar which has a nutrient content to promote shoot growth = needs high concentration of auxins compared to cytokinins
- Once the shoot grows move it to a new agar which has nutrient content to promote root growth = needs low concentration of auxins compared to cytokinins = gives rise to tiny plants called plantlets.
- Move plantlets to a greenhouse to be grown in soil/compost

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21
Q

Advantages of artificial cloning in plants

A
  • Fast
  • Can be done when sexual reproduction is not possible
  • Plant would have genetically identical features to parents = if these are desirable characteristics e.g. pest resistant or disease resistant, this can be beneficial
  • Plants have the same phenotype = easier to grow + harvest them together
  • When using meristems = new plants are free from viruses
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22
Q

Disadvantages of artificial cloning in plants

A
  • labour intensive
  • Expensive to set up the ideal conditions = specialised equipment
  • Low genetic diversity = all clones are susceptible to same diseases
  • No genetic diversity unless there’s a mutation
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23
Q

Use of reproductive cloning

A

for selective breeding, to increase the yield of a good characteristic

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24
Q

2 main techniques of reproductive cloning

A

Embryo twinning and somatic cell nuclear transfer

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25
Q

Embryo twinning

A
  • fertilise the egg with a sperm in vitro of two high value animals of the same species
  • Let zygote divide by mitosis to form small balls of cells
  • Impregnate surrogate mothers with small balls of cells
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26
Q

Somatic cell nuclear transfer

A
  • Take body cells from the sheep you want to clone and take an egg donor
  • Remove the nucleus from both the egg and the body cell and place the desired nucleus into the empty egg cell
  • Fuse them together using electrofusion
  • Put the new egg cell into the sheep so that it starts replicating and collect the early embryo
  • Implant the early embryo into a surrogate mother
  • This clones the exact body cell’s phenotype
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27
Q

non reproductive cloning in animals

A
  • process of cloning cells/tissues but not the whole animal
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28
Q

What can you do with non reproductive cloning in animals

A
  • Can grow skin in a lab = can be used to repair damage
  • Can be used for medical damage e.g. spinal chord issues, restoring the function of the pancreas
  • Can grow organs in the lab
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29
Q

Benefit of non reproductive cloning in animals

A

less chance of rejection bc it’s the patient’s own body cells

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30
Q

Advantages of non reproductive cloning in animals

A
  • can produce high yield of ideal characteristics
  • Genetically identical embryos can be used for scientific research to check the effect of environment on a gene
  • Drugs can be tested on cloned cells and tissues to avoid testing on humans/animals
  • Therapeutic cloning reduces the chances of rejection
  • Individuals from endangered species can be cloned to increase their numbers
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31
Q

Disadvantages of non reproductive cloning in animals

A
  • lack of genetic variation = all susceptible to the same disease
  • Success rate is poor and the method is very expensive
  • Ethical issues regarding how long the embryo survives
  • Doesn’t increase genetic diversity
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32
Q

Biotechnology

A
  • use of microbes/ any living thing for commercial purposes
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33
Q

Most common uses of biotechnology

A

for food and job production

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34
Q

Why are microbes used in biotechnology

A

fast growth rate, no growth requirements

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35
Q

Examples of microbes in biotechnology (food)

A

Organism used:
- yeast to make ethanol for beer
- yeast as source of carbon dioxide for baking
- lactobacillus to make lactic acid for yoghurt + cheese
- mycoprotein as a source of protein for vegetarians

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36
Q

What is mycoprotein made from

A

Fungus

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37
Q

Microbes in Biotech examples (pharmaceutical drugs)

A

penicillium fungus for penicillin, other antibiotics and insulin

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38
Q

Microbes in Biotech examples (enzymes)

A

protease and lipase in washing powders uses enzymes taken from bacteria
lactase from bacteria to make lactose free milk

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39
Q

Bioremediation

A

bacteria and fungi can be used to cleanse water

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40
Q

ADVANTAGES OF USING MICROBES IN BIOTECH:

A
  • cheap + easy to grow
  • Fast reproduction rate
  • Production takes place at lower temperatures and pressures than normally required
  • Production isn’t climate dependent = can happen anywhere in the world
  • Can use byproducts from other reaction as nutrients for bacteria and fungus in these reactions
  • Easier to genetically modify microbes
  • Few ethical considerations
  • Easier to purify + isolate products for microbes than in the conventional method
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41
Q

Other forms of biotechnology

A
  • gene technology
  • Genetic modification + gene therapy
  • Selective breeding
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42
Q

Examples of using biotech to make food

A

Brewing, baking, cheese, yoghurt

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43
Q

What is brewing and what does it use and why

A

alcohol production + uses yeast = promotes anaerobic respiration in yeast. Grapes used bc grapes have yeast on their surface + also contains glucose and fructose

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44
Q

Explain brewing using grapes

A

When the grapes are crushed, the Yeast on surface uses sugars from the grapes to anaerobically respire to produce carbon dioxide and alcohol

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45
Q

Explain brewing for beer

A

use barley grains just at the peak of germination = as the grains start to germinate they convert starch into maltose = used by yeast to respire anaerobically to produce co2 and alcohol.

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46
Q

Baking

A

yeast used = proof dough to allow the yeast to respire anaerobically = produces the co2 that allows the dough to rise.

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47
Q

Cheese

A

milk is treated w bacteria that uses lactose to make lactic acid = acidifies the milk. Milk is then mixed with rennet which contains the enzyme rennin. Rennin coagulates milk under the presence of calcium ions= forms curd which is separated from the liquid components of milk. The bacteria keeps forming more lactic acid. Take the curd and press it into moulds = produces cheese. Microbes can also be used to flavour cheese.

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48
Q

Whic bacteria ferment milk for yoghurt production

A

lactobacillus and streptococcus thermophilus

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49
Q

Yoghurt production

A

Allow bacteria to ferment milk
Lactose converted to lactic acid
Denatures the milk proteins which allows the milk to coagulate. Bacteria partially digest the milk = easier to digest yoghurt. Fermentation process is what gives rise to the taste of yoghurt. Probiotics can be added to yoghurt from other microbes to make it more nutritionally beneficial.

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50
Q

What else can biotechnology be used to make

A

Penicillin and insulin

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51
Q

Biotech for penicillin

A

secondary metabolite of the fungus penicillium

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52
Q

Biotech for insulin

A

genetically modify E. coli bacteria to produce human insulin

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53
Q

Bioremediation

A
  • when microbes are used to clean soils and underground water
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54
Q

What does bioremediation do

A

Convert toxic pollutants to less harmful substances

55
Q

Example of bioremediation

A

pseudomonas = treats oil spills = need to create the right conditions for the microbes to use the pollutants as a food source.

56
Q

How do you create the right conditions for the microbes to use the pollutants as a food source

A

by changing the temperature, the pH, adding additional nutrients.

57
Q

Advantages of bioremediation

A
  • use a natural system
  • less equipments used
  • fewer waste products
  • less risk of exposure to the workers
  • treatment happens in situ
58
Q

Is bio remediation done in situ or ex situ

A

can be done in situ, ex situ depending on if the conditions are favourable in situ

59
Q

Quorn

A

Aka single celled proteins

fungus produces single celled proteins e.g. mycoprotein acts as meat substitute.

60
Q

Advantages of using fungal protein (quorn)

A
  • faster to use plant (fungus) proteins
  • Production can either increase or decrease to demand
  • No animal killing
  • No animal fat or cholesterol
  • Can genetically modify proteins to increase it’s content of amino acids
  • Not much land is required
61
Q

Disadvantages of using fungal protein (quorn)

A
  • some people don’t want to eat fungal proteins
  • Isolation process needed to separate protein from fungus = has to be purified = extra step
  • Can cause infections
  • Although it’s a good source of protein, it’s amino acid profile will be different to that of protein
62
Q

What is used for the mass production of quorn

A

Fermenters

63
Q

Why is a fermenter used for mass production of quorn

A
  • Can control its conditions e.g. temperature, nutrient availability, oxygen availability, concentration of product
64
Q

What’s the most important step for mass production of quorn

A

to sterilise the fermenter using a rlly high temperature initially.

65
Q

What else is used in the mass production of quorn and why

A
  • pH probe = monitor pH to maintain optimal conditions for the growth of whatever microorganisms you want
  • Water jacket = temperature control
  • Air inlets = to provide the right amount of oxygen
  • Inlets for nutrients too
66
Q

2 types of culture

A

Batch and continuous

67
Q

Continuous culture

A

continuously promote the growth of your product = keep adding nutrients + remove some product + keep adding nutrients. Remove product bc product build up could be toxic.

68
Q

Batch culture

A

culture set up with limited quantity of nutrients + allow it to ferment for a while and then take out the product after the set time

69
Q

Why is using batch and continuous cultures important

A

bc microbes produce 2 types of products, primary metabolites and secondary metabolites

70
Q

Primary metabolites

A

products that are released as part of the normal growth of a microbe = nothing special

71
Q

Secondary metabolites

A

released in cases of threat/selection pressure to compete w other microbes e.g. antimicrobials and antibiotics

72
Q

Importance of asepsis using fermenters

A
  • provide a nutrient medium = could also promote the growth of unwanted microorganisms so then you’ll have the ones you want to grow and their competitors competing for nutrients and space = will have a low yield of ur product and a contaminated product + competitors may also start producing toxic products. THIS IS WHY STERILE CONDITIONS ARE IMPORTANT.
73
Q

Why do you use a microorganism culture for viruses

A
  • Can’t grow viruses in a lab because they need the machineries of a host cell = take a sample and put it through PCR
74
Q

How are bacteria and fungus taken for microorganism cultures

A

Culture a swab

75
Q

Two types of growth medium for microorganism cultures

A

can grow bacteria and fungus or an agar plate or nutrient broth (liquid medium)

76
Q

Purpose of microorganism cultures having two growth mediums

A

they have nutrients that promote growth e.g. salts, sugars, some can have blood

77
Q

Why are aseptic techniques

A

To prevent contamination

78
Q

Aseptic techniques for microorganism cultures

A
  1. Wash your hands.
  2. Disinfect the working area.
  3. Have a Bunsen burner operating nearby to heat the air. This causes the air to rise and prevents air-borne microorganisms settling, It also creates an area around it of sterile air in which the microbiologist can work.
  4. As you open a vessel, pass the neck of the bottle over the flame to prevent bacteria in the air entering the bottie. The bottle should also be flamed as it is closed.
  5. Do not lift the lid of the Petri dish off completely - just open it enough to allow introduction of the desired microorganism.
  6. Any glassware or metal equipment should also be passed through the flame before and after contact with the desired microorganisms
79
Q

How to sterilise microorganism cultures

A

Put the Petri dish and the inoculating loop through an autoclave at 121 degrees celsius to sterilise everything.

80
Q

How to create agar

A
  • open bottle over Bunsen burner flame
  • When pouring agar gel and only open lid slightly, enough to pour the liquid into the plate
81
Q

Inoculation

A

introduction of microorganisms to the sterile medium (taking bacteria from a liquid broth and spreading it over agar)

82
Q

Different ways to inoculate?

A

Streaking, seeding, spreading

83
Q

Streaking

A

bc broth has lots of bacteria inside, when performing streaking the aim is to isolate one bacterial colony to see its morphology

84
Q

Difference between streaking and other techniques

A

Streaking isolates colony, other techniques allow you to grow a bacteria

85
Q

Steps of streaking

A
86
Q

Why is the agar lid slightly open

A

To prevent contamination

87
Q

Seeding

A

a sterile pipette is dipped in the liquid broth and this is dropped over the surface of the agar

88
Q

Spreading

A

using a sterile glass spreader to spread the bacteria from the liquid broth over the surface of the agar

89
Q

What else can be used for inoculation instead of seeding or spreading

A

Can also use a moist cotton swab with distilled water and placing this over the agar and closing the agar

90
Q

Incubation

A

keeping the taped up agar plate at a specific temperature for a set amount of time.

91
Q

What should you do when closing the lid for incubation

A

Don’t tape it fully

some air movement to allow for the growth of some organisms. And make sure the Petri dish is labelled.

92
Q

What should you do when storing the Petri dish for incubation

A

store it in a warm environment to allow for bacterial growth but store it upside down = prevents condensation from falling back onto the microorganisms

93
Q

What do bacteria and fungus look like after an incubator is used

A

If it’s spongey + green = likely to be a fungus
Shiny circles = bacteria

94
Q

Using a liquid medium

A

allows bacteria to grow but you won’t be able to identify characteristics of the bacteria but can measure population growth by seeing how much light passes through the broth

95
Q

Serial dilution using streak plating

A

If there’s x amount in the smallest dilution you can work your way backwards to calculate how many were in the original broth culture.
= use serial dilution and plating.

96
Q

Serial dilution using streak plating diagram

A
97
Q

Units for counting bacteria

A

cfu (colony forming units)

98
Q

Population growth curve in a closed culture

A
99
Q

Lag phase

A

time taken for bacteria to acclimatise to the environment. E.g. cell growth, switching on genes, protein synthesis.

100
Q

Exponential phase

A

when the bacteria are used to environment and start reproducing and there’s plenty of nutrients and space = no setbacks

101
Q

Stationary phase

A

when the organisms have started to use up the nutrients and there’s more waste in the system now and the reproduction rate and death rate are equal

102
Q

Death/decline phase

A

when organisms run out of nutrients and there’s a build up of waste so some organisms are killed.

103
Q

When are primary metabolites made in terms of the population growth curve

A

made part of the natural growth and are normally made during the log phase

104
Q

When are secondary metabolites made in terms of the population growth curve

A

only produced when the bacteria sense danger (when it wants to start competing with someone) e.g. antibiotics = produced late stationary phase/early decline phase.

105
Q

Are antibiotics primary or secondary metabolites?

A

Secondary

106
Q

How do you collect a secondary metabolite

A
  • create a closed system w limited resources = induces stationary/decline phase environment
107
Q

Immobilised enzymes

A
  • new invention where enzymes are held in place and aren’t free in solution = can immobilise them to surfaces where they’re stuck to something.
108
Q

Advantages of immobilising enzymes

A
  • if you wanted to make a solution and needed an enzyme just pass a substrate through a mixture and the product will gather at the bottom.
  • Enzymes aren’t used up in the reaction but they remain in the beaker = means at the end they’d need a purification step
  • Benefits of this is that enzymes don’t mix with the products, so the purification step would be cheaper
  • Easier to reuse the enzymes
  • Only putting enzymes in rather than cells that make the enzymes = better bc all the requirements for the cell e.g. nutrients and reproduction stages would no longer be needed
  • By immobilising the enzymes you are protecting the enzymes from extreme conditions = high temperature and pH can be used.
109
Q

Methods for enzymes to bind to surface

A

Adsorption, covalent bonding, entrapment, entrapment w membrane separation

110
Q

Adsorption

A
  • Bind enzymes to a surface using hydrophobic interactions and ionic links.
  • Aim is to stick the back of the enzymes to these surfaces so that the active site is exposed but because using ionic links, the shape of the active site might get slightly distorted.
111
Q

What can enzymes bind to in adsorption

A

Clay, glass beads, resins

112
Q

How can enzymes detach in adsorption

A

can detach from the surface because the hydrophobic interactions and ionic links are weak.

113
Q

Covalent bonding

A
  • enzymes use covalent bonds to bind to the clay particle and cross linking agents can be used to add more enzymes
114
Q

Benefit and negative of covalent bonding

A

covalent bonds are strong so they are unlikely to come out. But because the bonds are strong they are more likely to distort the active site shape = reducing activity

115
Q

Entrapment

A
  • trap the enzyme in a matrix = enzymes aren’t directly exposed to the substrate = substrate and products have to go through the matrix and then come out of the matrix.
116
Q

Why is entrapment safer for enzymes

A

because there’s no bonding involved which can alter the active site = only enclosed in matrix but the products and substrate would have to diffuse out of the matrix so they’d have to be smaller molecules = not suitable for all reactions.

117
Q

Example of entrapment medium

A

Cellulose mesh

118
Q

Entrapment a membrane separation

A
  • membrane separating the enzymes and the product and substrate = the product and substrate will only pass through if they’re permeable to the membrane. = alternative version of entrapment
119
Q

Industrial uses of immobilised enzymes

A

Glucose isomerase, penicillin acylase, lactase, aminoacyclase, glucoamylase

120
Q

Glucose isomerase

A
  • converts glucose to fructose, which is now used to make most of the sugary food because it a lot sweeter so you need less sugar
121
Q

Penicillin acylase

A
  • Form semi synthetic penicillin = to get ahead of antibiotic resistance
122
Q

Lactase

A
  • breaks down lactose to glucose and galactose
  • Produce lactose free milk
123
Q

Aminoacyclase

A
  • Produces pure samples of L amino acids = can be used in the pharmaceutical industry to create compounds
124
Q

Glucoamylase

A
  • converts dextrins to glucose
125
Q

What are viruses

A

Bits of genetic code

126
Q

Do viruses have dna or rna

A

Some have dna, some have rna

127
Q

Examples of viruses w rna

A

HIV, influenza and rabies

128
Q

Examples of viruses w dna

A

Warts and chickenpox

129
Q

Which virus is most likely to mutate + why

A

RNA viruses = more likely to mutate because there’s only one strand = risked

130
Q

What do viruses rely on

A

Reverse transcriptase

131
Q

What’s a virus’s aim

A

To get it’s genetic material inside a cell

132
Q

Disadvantages of using microorganisms

A

Some people may not want to eat fungal protein or food grown on waste
• Need to isolate the protein from the medium it is grown in
• Protein has to be purified
• Culture can become infected easily due to optimum growing conditions
• Different taste/texture to traditional protein sources

133
Q

Why can no population grow indefinitely

A

Limiting factors e.g.
Food
• Shelter
• Light
• Water
• Oxygen
• Accumulation of toxic waste
• Disease
• Predators

Biology Module 6 (6 decks)

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