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Biology - Module 6 > Cloning and Biotech > Flashcards

Cloning and Biotech Flashcards

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1
Q

vegetative propagation / natural cloning

A

reproduce asexually using meristem cells

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2
Q

what do you need for vegetative propagation

A

propagate asexually using tubers, rhizomes, bulbs, suckers, and offsets

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3
Q

Rhizome

A

specialised horizontal stem running underground + stores food – buds develop

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4
Q

Vegetative organs of plants

A

enable plants to survive in adverse conditions – contain food + remain dormant

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5
Q

examples of vegetative organs of plants

A

Root and shoot tips

Axillary buds (where leaves and the stem meet)

Vascular cambium (between xylem and phloem)

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6
Q

how does natural cloning take place

A

over time miniature plant (a plantlet) / buds forms at these locations + remains attached to its parent plant

clones of their parent

At maturity = detaches

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7
Q

potatoes

A

Potato tubers are swollen modified roots that form eyes on their surface

Eyes can sprout new growth (called ‘chitting’)

The starch stored in the tuber fuels the early growth of the new plant

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8
Q

ginger

A

Ginger forms rhizomes, a modified stem that grows horizontally underground

New growth stems from nodes in the rhizome, forming new stems and adventitious roots

The section used in cookery is the rhizome

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9
Q

strawberries / spider plants

A

have horizontal stems or runners that form over the soil surface, pointing sufficiently far away

new plant = not be overshadowed by its parent, or in competition for water or soil nutrients

Roots form under the nodes of runners, called adventitious roots

The runner dies when the plantlet is self-sustaining

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10
Q

state the type of plant tissue in which clones are produced

A

meristematic

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11
Q

State methods of natural cloning in plant

A

runners / suckers / stolons / tubers /
rhizomes / bulbs

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12
Q

how to propagate from cuttings

A

Short sections of stems taken + planted directly on ground or in pots

cut stems at a slant between nodes

Rooting hormones applied to base of cutting – encourage growth of new roots

remove leaves - reduce transpiration

natural clone

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13
Q

Cuttings vs seeds

A

Faster

Guarantees quality of plants

Lack of genetic variation

Sugar canes

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14
Q

factors that increase success rate in cuttings

A
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15
Q

how does using non flowering plant increase success rate

A

all plant resources available for growing new roots

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16
Q

how does making an oblique cut increase success rate

A

maximises surface area available for rooting powder/new root development

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17
Q

how does using a hormone rooting powder increase success rate

A

scientists unsure whether effect is the hormone directly or anti-fungal action but seems to increase success rate

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18
Q

how does reducing leaves increase success rate

A

minimises loss of water by transpiration whilst maintaining photosynthesis

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19
Q

how does keep cutting well watered increase success rate

A

reduces water stress

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20
Q

how does covering cutting with a plastic bag increase success rate

A

keeps air humid and reduces water loss by transpiration

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21
Q

Micropropagation

A

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

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22
Q

why does micropropagation work

A

Plant cells – totipotent – entire plant can be reproduced from any of these cells

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23
Q

process of micropropagation

A

Take small sample of tissue from plant

Sample sterilised by immersing it in sterilising agents

Material removed from plant – explant

Explant placed in sterile culture medium

Balance of plant hormones – e.g auxins + cytokinins – simulate mitosis

Cells proliferate – forms callus = mass of identical cells

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24
Q

where should we take a small sample of tissue from in micropropgation

A

Meristem tissue from shoot tips + apical buds

Sterile conditions – virus-free

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25
Q

sterilising agents

A

bleach / ethanol / sodium dichloroioscyanurate

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26
Q

explant

A

Material removed from plant

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27
Q

Advantages of plant cloning

A

same genotype + phenotype

The plants produced are free of disease

genetically modified to confer immunity to certain diseases

rapid and can yield large numbers of new plants

Plants that are difficult to grow from their seeds can be produced by plant cloning

Plants can be grown in any country, in any season

Rare and endangered species can be propagated to save them from extinction

Whole plants can be created from genetically modified cells/tissues

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28
Q

Disadvantages of plant cloning

A

expensive and labour-intensive process

susceptible to microbial contamination

no genetic variation, so all of the offspring are susceptible to the same diseases or other environmental factors

risks large-scale loss of a country’s / continent’s crop of a particular plant

plants have to be carefully screened for abnormalities that could lead to the new plants being infected

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29
Q

natural clones in animals

A
  • invertebrates - regenerate animals from fragments of original

monozygotic twins

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30
Q

why identical twins are referred to as monozygotic

A

from the same zygote

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31
Q

how are twins formed

A

egg fertilised by a sperm

forms a zygote

single zygote undergoes a few cell cycles = embryo

embryo splits in two

Two embryos that form are identical

identical offspring, always of the same gender, with identical phenotype

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32
Q

why are identical twins not clones

A

mutations occur in every cell cycle

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33
Q

why may identical twins look different when born

A

difference in position + nutrition in uterus

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34
Q

artificial clones in animals

A

artificial twinning + somatic cell nuclear transfer

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35
Q

artificial twinning principle

A

artificially split embryo - can be split into more than 2

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36
Q

artificial twinning process

A

cows – treated with hormones – super ovulates

ova fertilised naturally or via artificial insemination

early embryos flushed out of uterus

OR – eggs fertilised in lab

Before day 6 – cells still totipotent

Cells of embryo split

Grown in the lab for a few days

Implanted into surrogate mother – each different mother for cows– single pregnancies carry less risk

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37
Q

Somatic cell nuclear transfer / reproduction cloning

A

Nucleus removed from somatic cell of an adult animal

Nucleus removed from a mature ovum harvested from different female animal of the same species

Enucleated ovum

Nucleus from adult somatic cell placed into enucleated ovum

Milk electric shock – fuses + begins to divide

OR – nucleus from adult cell not removed + placed next to enucleated ovum – divide due to electrofusion – electric current

Embryo – put into uterus of a third animal

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38
Q

DNA of offspring from SCNT

A

clone of the animal from which the original somatic cell is derived

BUT mitochondrial DNA – come from egg cell

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39
Q

animals required for SCNT

A

3

The animal to be cloned by donating a cell

The female to donate an egg cell

The surrogate mother

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40
Q

SCNT vs artificial twinning

A

Artificial twinning – clones embryo

SCNT – clones adult animal

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41
Q

Arguments for animal cloning

A

Artificial twinning – high yielding farm animals to produce more offspring

Enables success of male animal at passing on desirable genes to be determined

SCNT enables GM embryos to replicate / develop – important in pharming

SCNT – enable rare / endangered / extinct animals to be reproduced

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42
Q

Arguments against animal cloning

A

SCNT – inefficient – takes many eggs to produce single cloned offspring

Cloned embryos – fail to develop / miscarry / malformed offspring

Animals produced via cloning – shortened lifespans

Cloning destroys embryos which could in theory develop into a healthy adult animal

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43
Q

Biotech

A

applying biological organisms / enzymes to the synthesis / breakdown / transformation of materials in the service of people

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44
Q

why are microorganisms ideal for biotech

A

No welfare issues to consider – only optimum conditions for growth

Enormous range of microorganisms capable of carrying of chemical syntheses / degradations

GM – manipulate microorganisms

Short life cycle + rapid growth rate

Nutrient requirements- simple + cheap

Occupy very little space

45
Q

microorganisms + purpose in baking

A

yeast

mixed with sugar to respire aerobically

carbon dioxide produced makes bread rise

46
Q

steps in commercial process - baking

A
47
Q

microorganisms + purpose in brewing

A

yeast

respires anaerobically to form ethanol

GM yeasts ferment at lower + cheaper temp

48
Q

steps in commercial process - brewing

A
49
Q

microorganisms + purpose in cheese making

A

bacteria

feed off lactose in milk - changing texture + tase

inhibiting growth of bacteria that make milk go off

50
Q

steps in commercial process - cheese making

A
51
Q

microorganisms + purpose in yoghurt making

A

bacteria

forms ethanal + lactic acid

extracelular polymers that give yoghurt smooth thick texture

52
Q

steps in commercial process - yoghurt making

A
53
Q

advantages of using microoroganisms for food

A
54
Q

disadvantages of using microoroganisms for food

A
55
Q

making penicillin

A

P. chryogenum – requires high oxygen levels + rich nutrient medium to grow well

Semi-continuous batch process used

Species of mould from the Penicillium genus can be cultured in industrial fermenters

deep-tank fermentation

Extraction and purification of the product produces large volumes of the drug for therapeutic use

56
Q

making penicillin conditions

A
57
Q

making insulin

A

Bacteria grown in fermenter + downstream processing results in constant supply of pure human insulin

Recombinant DNA technology can incorporate the gene for human insulin into the genome of the bacterium, Escheriscia coli

Recombinant bacteria are grown in batch fermenters, and each bacterial cell expresses insulin

Insulin is released into the batch medium and purified

58
Q

Bioremediation

A

Microorganisms used to break down pollutants + contaminants in soil / water

Naturally occurring microorganisms perform aerobic digestion of the contaminants and release non-polluting products

59
Q

how does bioremediation work - natural vs GM

A

Use natural organisms

Many microoganisms naturally break down organic material – CO2 + water

Break down + neutralise contaminants

Oil spill – add nutrients – encourage microbial growth - biostimulation

GM organisms

Break down / accumulate contaminants that they don’t usually encounter

60
Q

biostimulation

A

add nutrients – encourage microbial growth

61
Q

bioventing

A

process which allows oxygen to reach the contaminants

62
Q

what does bioremediation rely on

A

oxidative digestion of pollutants

Naturally occurring microorganisms perform aerobic digestion of the contaminants and release non-polluting products

63
Q

why do you need to be careful even with harmless microorganisms

A

Risk of mutation + becoming pathogenic

Contamination with pathogenic microorganisms from environment

64
Q

nutrient medium

A

Food provided to bacteria for culturing

liquid form - broth
solid form - agar

65
Q

Inoculating broth

A

Make suspension of bacteria to be grown

Mix known volume with sterile nutrient broth in flask

Stopper the flask with cotton wool – prevent contamination

Incubate at temp – shake regularly to aerate the broth – provide oxygen for bacteria

66
Q

Inoculating agar

A

Wire inoculating loop – sterilised by holding in Bunsen flame until it grows red hot

Must not be allowed to touch any surfaces as it cools – avoid contamination

Flame the neck of the culture tube

Dip sterilised loop into bacterial suspension in culture tube

Remove lid of petri dish + make a zig-zag streak across surface of agar

Avoid digging loop into agar

Replace the lid of petri dish – held down with tape but not sealed completely

Oxygen can still get in – prevent anaerobic bacteria

Incubate

67
Q

general aseptic techniques include:

A

Washing hands thoroughly

No food or drink allowed in the lab

Disinfecting work surfaces with disinfectant or alcohol

Wearing gloves and goggles

Working close to a lit Bunsen burner

Flaming equipment (to kill microorganisms or create updraughts)

Sterilising (in an autoclave) or disposing of all used equipment

68
Q

why should we work close to a bunsen burner

A

(this creates an updraught of air so prevents contamination from airborne fungal spores, for example)
kill micro-organisms

69
Q

purpose of these steps

A
70
Q

Primary metabolites

A

substances formed as an essential part of the normal functioning of microorganism

E.g. ethanol

71
Q

Secondary metabolites

A

substances produced that are not essential for normal growth but still used by cell

E.g. pigments

72
Q

significance of primary + secondary metabolites

A

depending on which you want - determine the time in which you will harvest the culture

73
Q

Batch fermentation

A

Microorganism inoculated onto fixed volume of medium

Growth takes place

Nutrients used up

New biomass + waste products build up

Culture reaches stationary phase – overall growth stops

Often carry our changes to make desired products

Process stopped before death phase + products harvested

74
Q

Continuous fermentation

A

Microorganisms inoculated into sterile nutrient medium

Medium added continually to culture once it reaches exponential point of growth

Culture broth continually removed – medium / waste products / microorganisms / product

Keep culture volume in bioreactor constant

75
Q

Downstream processing

A

Bioreactors – produce mixture of unused nutrient broth / microorganisms / primary + secondary metabolites / waste

Useful part has to be separated – downstream processing

76
Q

factors that will max yield of products in bioreactors

A

temp

nutrients

oxygen

mixing

asepsis

77
Q

controlling temp

A

Maintain optimum temp – rate + denature

Heating / cooling systems linked to temp sensors

Negative feedback system

E.g. – use water jacket

Max enzyme activity = max yield

78
Q

controlling nutrients

A

Added + circulated to ensure access

Probes / sample tests indicate levels dropping

79
Q

controlling oxygen

A

Sterile air pumped in

Provided max oxygen – max respiration – max yield

80
Q

mixing

A

Large volumes of liquid – viscous

Simple diffusion not enough to ensure food + correct temp

Mixing mechanisms – stirred continuously with paddles

Even distribution

81
Q

asepsis

A

Sealed / aseptic units

Must be cleaned between cultures – prevent contamination

Contamination – interspecific competition – reduce yield

82
Q

state the growth phases of bacterial colonies in a closed system

A

lag

log

stationary

death

83
Q

describe the growth of bacterial colonies in a closed system

A
84
Q

measuring bacterial populations

A

Direct counting – includes all cells – living or dead

Viable counting – culturing samples + counting colonies that grow – only takes living samples into account

Turbidity – measure of living + dead in solution

85
Q

turbidity

A

Grow in broth

Turbidity – measure of cloudiness of a suspension

As population grows – becomes more turbid

changing turbidity - monitored by measuring how much light can pass through the suspension at fixed time intervals after the initial inoculation – colorimeter – plot a curve

86
Q

why log phase

A

high availability of nutrients + plenty of space

87
Q

why death phase

A

due to lack of nutrients + toxic wase builds up

88
Q

how to calculate the number of bacteria after divisions

A
89
Q

factors that limit the log phase

A

nutrients - as bacteria multiply - nutrients used up + becomes insufficient to support growth

oxygen - as population increases - demands for respiratory oxygen increases

temp - too low / too high

build up of waste - toxic material inhibit growth + kill culture

change in pH - carbon dioxide produced by respiration = pH falls - effects enzyme activity

90
Q

advantages of isolated enzymes over whole organism

A

less wasteful – whole microorganism use up substrate growing + reproducing – makes biomass

more efficient – isolated work at much higher conc

more specific – no unwanted enzymes + no wasteful wide reactions

less downstream processing – pure product produced // whole organisms produce variety of products difficult + expensive to purify

91
Q

immobilised enzyme

A

enzyme that is attached to an insoluble material to prevent mixing with the product

92
Q

advantages of immobilised enzymes

A

held stationary during reactions – can be recovered from mixture + reused

enzymes do no contaminate end product - no downstream processing

greater temp tolerance – less easily denatured by heat – optimum over a much wider temp range – bioreactor less expensive to run

93
Q

disadvantages of immobilised enzymes

A

Specialist expensive equipment required – high cost of bioreactor

more costly to buy - unlikely to be financially worthwhile for smaller industries // higher initial cost of materials

rate of reaction is sometimes lower - as the enzymes cannot freely mix with the substrate

94
Q

4 ways of immobilising enzymes

A
95
Q

adsorption

A

surface immobilisation

adsorbed to inorganic carriers - cellulose / silica / carbon nanotubes

96
Q

advantages + disadvantages of adsorption

A
97
Q

covalent / ionic bonding

A

surface immobilisation

covalent bonding - carriers with amino / hydroxyl /carboxyl groups

ionic bonding - polysaccharides - cellulose

98
Q

advantages + disadvantages of covalent / ionic bonds

A
99
Q

entrapment - in matrix

A

polysaccharides / gelatin

100
Q

advantages + disadvantages of entrapment in matrix

A
101
Q

entrapment - capsules

A

membrane entrapment in microcapsules

encapsulation

102
Q

advantages + disadvantages of entrapment - encapsulation

A
103
Q

example of immobilised enzymes forming lactose free dairy products

A

Enzyme: Lactase

Converts lactose to glucose and galactose

104
Q

example of immobilised enzymes forming Semi-synthetic penicillin

A

Enzyme: Penicillin acylase

Converts the original form of penicillin into one which is effective against penicillin-resistant organisms

105
Q

example of immobilised enzymes forming sweetened / thickened food

A

Enzyme: Glucoamylase

Converts starch and other dextrins into glucose

106
Q

example of immobilised enzymes forming sweetened foods with low sugar

A

Enzyme: Glucose isomerase

Converts glucose into the sweeter sugar, fructose

106
Q

example of immobilised enzymes forming purified samples of L amino acids

A

Enzyme: Aminoacylase

Separates out L-amino acids from D-amino acids

107
Q
A

Biology - Module 6 (6 decks)

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