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A Level Biology > Cloning and Biotechnology > Flashcards

Cloning and Biotechnology Flashcards

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1
Q

E.g. of immobilised enzymes: glucose isomerase

A

for the conversion of glucose to fructose

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2
Q

E.g. of immobilised enzymes: penicillinacyclase

A

acyclase for the formation of semi-synthetic penicillins (to which some penicillin-resistant organisms are not resistant)

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3
Q

E.g. of immobilised enzymes: lactase?

A

for the hydrolysis of lactose to glucose and galactose

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4
Q

E.g. of immobilised enzymes: aminoacyclase?

A

for production of pure samples of L-amino acids

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5
Q

E.g. of immobilised enzymes: glucoamylase?

A
  • for the conversion of dextrins to glucose
  • dextrins are a group of low-moelcular weight carbs produced by the hydrolysis of starch or glycogen
  • dextrins are mixtures of polymers of alpha glucose units linked by alpha1,4 or alpha1,6 glycosidic bonds
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6
Q

E.g. of immobilised enzymes: nitrilase?

A

for the conversion of acrylonitrile to acrylamide (for use in the plastics industry).

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7
Q

What is a clone?

A

genetically identical copy e.g. monozygotic twins

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8
Q

what is horticulture?

A

The growing of flowers, fruits, vegs on a relatively small scale - gardening. Contrasts w agriculture which is on a large scale

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9
Q

which organisms reproduce asexually?

A
  • plants
  • all prokaryotes
  • eukaryotes that reproduce by mitosis
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10
Q

advantages of of cloning?

A
  • quick - no pollination involved
  • can happen w/o mate
  • all offspring cope in that enivr
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11
Q

disadvantages of cloning?

A
  • no genetic variation
  • so any genetic weaknesses passed on
  • e.g. selection pressure - all would die
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12
Q

artificial cloning in plants ?

A
  • Gardners take cuttings of plants
  • can make use of the fact that plant vascular tissue can join up in the process of grafting
  • A scion is cut and grafted onto root stock
  • all grape vines are produced like this
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13
Q

natural cloning is a.k.a

A

vegetative propagation

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14
Q

natural cloning occurs in?

A
  • Bulbs e.g. daffodils
  • runners e.g. strawberry
  • stem tubes e.g. potato
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15
Q

using natural cloning in horticulture?

A
  • Possible to take cuttings of many plants - short sections of stems are taken and are planted either directly into the ground or in pots
  • Rooting powder often applied to the base of the cutting to encourage growth of new roots
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16
Q

Propagation from cuttings advantages over using seeds?

A
  • faster - time from planting to cropping is reduced

* guarantees quality of plants

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17
Q

Propagation from cuttings disad compared to using seeds?

A

lack of genetic variation

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18
Q

in natural cloning what forms?

A

a structure that develops into a fully differentiated new plant, which is gen identical to parent

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19
Q

2 basic ways of veg prop?

A
  • cuttings

* runners

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20
Q

⭐ many organisms can reproduce ? and

A

asexually and sexually

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21
Q

some plamnts can also reproduce sexually which…

A

leads to variation via seed dispersal. Seed can be carried to a diff area where selection pressures are diff which may lead to speciation.

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22
Q

Artificial cloning?

A
  • storing natural produce (tubers/runners/bulbs)
  • cuttings between lead nodes
  • grafting

e.g. commercial grape vines

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23
Q

`What is micropropagation?

A

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

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24
Q

What kind of tissue is used in micropropagation?

A

meristematic - totipotent - can differentiate to produce all the different types of cell in a plant

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25
Q

Plants that are cloned using tissue culture?

A
  • that don’t readily produce seeds
  • plants that are difficult to take cuttings from
  • rare and endangered plants
  • plants that have been GE
  • plants produced by selective breeding
  • plants that must be free of pathogens
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26
Q

Microprop a.k.a ?

A

tissue culture

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27
Q

Natural cloning in animals?

A
  • Many invertebrates capable - e.g. starfish, sponges
  • In vertebrates, monozygotic twins are formed from embryo splitting
  • Parthenogenesis
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28
Q

Parthenogenesis?

A

Natural form of asexual reproduction in which growth and development of embryos occurs w/o fertilisation. In animals means development of an embryo from an unfertilised egg cells

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29
Q

Artificial cloning 2 man techniques:

A
  • Artificial embryo twinning

* Somatic cell nuclear transfer

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30
Q

Embryo splitting?

A
  • early embryo from cow and bull with desirable trait split into totipotent cells
  • totipotent cell develops into an embryo
  • embryos transferred to surrogate mothers
  • identical cloned offspring produced
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31
Q

Somatic cell nuclear transfer?

A
  • Diploid cell taken from mammary gland
  • Nucleus removed from somatic cell
  • Egg cell taken from egg cell donor
  • Nucleus removed
  • Nucleus of somatic cell put into egg cell
  • Electric pulses used to fuse nucleus and egg cell, causing it to /
  • Cell /, forming embryo
  • embryo implanted into surrogate mother
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32
Q

offspring produced by somatic cell nuclear transfer?

A

genetically identical to nuclear donor, but mitochondrial DNA will come from egg cell

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33
Q

+ of SCNT?

A
  • genetically identical - more sheep w desirable characteristics
  • can be used to produce GMA which grow organs that can be used in human transplants
  • Allows GM embryos to be replicated and to develop, giving many embryos from 1 GE procedure
  • spec animals can be cloned
  • rare/ endangered species reproduced
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34
Q
  • of SCNT?
A
  • Lack of GV dec robustness of population
  • any genetic diseases, mutations passed down
  • premature ageing
  • inefficient - many eggs to produce a single cloned offspring
  • Many clones embryos fail to develop and miscarry
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35
Q

+ of embryo splitting?

A
  • more offspring produced than normal reproduction

* allows success of make animal at passing on desirable genes to be determined

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36
Q
  • of embryo splitting?
A
  • Many animals produced by cloning have red. life spans

* many cloned embryos fail to develop and miscarry or produce malformed offspring

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37
Q

Non - reproductive cloning?

A

• Cloning specific cells to replace others, not the whole animal`

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38
Q

Why clone cells?

A
  • testing to see effect of external factors on the cells
  • research effects of radiation
  • cosmetics (reduced need for animal testing)
  • now used experimentally for gene therapy
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39
Q

What is biotechnology?

A

using biological organisms/ enzymes for the synthesis, breakdown or transformation of materials for the benefit of humans
e.g. yeast - fermentation

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40
Q

Cheese?

A
  • ester bonds broken by enzymes in bacteria

* FA = acidic, causes milk to curdle

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41
Q

What is most commonly used in biotechnology?

A

Fungi e.g. yeast or bacteria e.g. E coli

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42
Q

Why are microorg used in biotechnology?

A
  • reproduce quickly - rapid growth and v short life cycle
  • cheap molecular req. - can often use the wate products of other industry
  • no ethical issues
  • convenient - no welfare issues
  • easier to store
  • wide range available
  • can be genetically modified - 1 chromosome in bacteria
  • grow in simple conditions - high T and P not needed
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43
Q

E.g. of bacteria used in biotech?

A

E. coli - produce GM insulin and lactase

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44
Q

E.g. of archae used in biotech?

A

Thermoccocus - heat resistant enz e.g. Taq polymerase

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45
Q

E.g. of fungi used in biotech?

A

Penicillium - penicillin

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46
Q

E.g. of protoctists used in biotech?

A

algae - food thickening agents

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47
Q

E.g of biotech processes: baking?

A
  • yeast
  • anaer to produce ethanol and CO2
  • CO2 makes bread rise
  • yeast added to flour, left in warm envir to rise
  • excess air removed, left to rise again
  • cooked in hot oven, CO2 bubbles expand so bread rises more
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48
Q

E.g. of biotech processes: brewing?

A
  • yeast, produced ethanol
  • enzymes hydrolyse starch to sugars that can be used by the yeast
  • enzymes break down starch to wort
  • wort sterilised and cooled
  • wort inoculated w yeast
  • beer is conditioned for 4-29 days at 2-6 in tanks
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49
Q

E.g. of biotech processes: cheese making?

A
  • bacteria feed on lactose in milk, changing texture and taste and inhibiting the growth of bacteria which make milk go off
  • Milk pasteurised and homogenised
  • mixed with bacterial cultures and kept until the milk separated into solid curds and liquid whey
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50
Q

E.g. of biotech processes: Yoghurt making?

A
  • Bacteria to produce ethanal and lactic acid
  • skimmed milk powder added to milk, mixture pasteurised, homogenised, cooled
  • milk mixed with a 1:1 ratio w lactobasillus bulgarius and streptococcus thermophilus and incubated at around 45 degrees for 4-5 hrs
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51
Q

E.g. of biotech processes: Penicillin ?

A
  • produced by the mould penicillium notatum
  • semi-conservative batch process used
  • first stage of production: fungus grows
  • 2nd stage: penicillin produced
  • then drug extracted and purified
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52
Q

E.g. of biotech processes: insulin?

A
  • less side effects
  • Used by more religious groups
  • Bacteria grown in a fermenter and downstream processing results in a constant supply of pure human insulin
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53
Q

E.g. of biotech processes: bioremediation?

A
  • MO used to break down pollutants and contaminants in the soil in water
  • natural organisms can be used - many MO naturally break down organic material producing CO2 +H2O
  • Or GM MO - scientists trying to develop GM bacteria which can break down or accumulate contaminants which they would not naturally encounter
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54
Q

Advantages of using MO to produce human food: fast

A

reproduce fast and produce proteins faster than plants and animals

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55
Q

Advantages of using MO to produce human food: WASTE

A

can be fed on waste, reducing costs

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56
Q

Advantages of using MO to produce human food: PROTEINS

A

MO can be genetically modified to produce req proteins

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57
Q

Advantages of using MO to produce human food: welfare issues

A

no welfare issues w growing MO

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58
Q

Advantages of using MO to produce human food: not dependent on

A
  • production of MO is not dependent on weather, breeding cycles etc
  • takes place constantly
  • and can be inc or dec to meet demand
59
Q

Advantages of using MO to produce human food: protein and fat content?

A

MO have a high protein content with little fat

60
Q

Advantages of using MO to produce human food: taste?

A

can be made to taste like anything

61
Q

Disadvantages of using MO to produce human food: toxins?

A

can produce toxins if conditions not maintained at optimum

62
Q

Disadvantages of using MO to produce human food: separated?

A

MO have to be sep. from nutrient broth and processed to make food

63
Q

Disadvantages of using MO to produce human food: sterile conditions?

A

need sterile conditions that are carefully controlled adding to costs

64
Q

Disadvantages of using MO to produce human food: GM organisms?

A

often involve GM organisms and many people have concerns about eating GM foods

65
Q

Disadvantages of using MO to produce human food: purification?

A

the protein has to be purified to ensure it contains no toxins or contaminants

66
Q

Disadvantages of using MO to produce human food: waste?

A

many ppl dislike the thought of MO grow on waste

67
Q

Disadvantages of using MO to produce human food: natural flavour?

A

little natural flavour - need additives

68
Q

What is culturing MO?

A

growing large pops of MO can be done using agar plates, broth or bioreactors

69
Q

Stages of a growth curve?

A
  1. the lag phase
  2. exponential phase
  3. stationary phase
  4. death phase
70
Q

The lag phase?

A
  • no. slowly inc (this is b4 growth is detected)

* bc adjusting to envir - synthesising enzymes needed e.g. lac operon

71
Q

Exponential phase?

A

• growth taking place at the max rate (doubling)

72
Q

stationary phase?

A
  • death rate = growth rate

* decreased availability of space, nutrients running out

73
Q

death phase?

A
  • death rate . growth rate
  • nutrients ran out
  • toxic build up
74
Q

when does a sigmoidal growth curve occur?

A

always happens when MO are grown in a culture where the nutrients are limited

75
Q

but if nutrients could be kept from running or what would happen?

A

stationary phase wouldn’t be reached, would stay in exponential phase

76
Q

equation to work out pop sizes?

A

• Nt = No x 2^n
Nt = no. of cells at end
No = no. of cells at start
• n = no. of generations

77
Q

why is 2 used?

A

pop doubles every generation

78
Q

if want to work out no. of generations?

A

use log little 2 (…)

79
Q

how many mm3 per c3?

A

1000

80
Q

how many cm3 per dm3?

A

1000

81
Q

What is a metabolite ?

A

• a molecule made by a cell in the course of its metabolism

82
Q

primary metabolite?

A
  • molecule produced by all of the cells in a culture all of the time
  • involved in normal metabolic processes like resp and growth
83
Q

secondary metabolite?

A

only produced by some cells or at a particular growth phase. Not directly involved in normal metabolic processes

84
Q

e.g. of primary metaboloute?

A
  • pyruvate produced in glycolysis

* urea by ornithine cycle

85
Q

secondary metabolite e.g.

A

penicillin produced by Penicillium fungus and Morphine produced by the opium poppy

86
Q

Batch culture?

A
  • MO inoculated into a fixed V of medium
  • As the culture is abut to reach the death phase the products are harvested
  • the whole system is then cleaned and sterilised and new batch set up
87
Q

Product of batch culture?

A

The product of a batch culture is often a secondary metabolite, produced in the stationary phase, when the population is under environmental stress.

88
Q

continuous culture?

A

• MO are inoculated into sterile medium and start to grow
• sterile nutrient medium is added continually to the culture once it reaches the exponential point of growth
• culture broth is continually removed - the medium waste products, MO, products
- keeping the culture volume constant

89
Q

Product of continuous culture?

A

The product of a continuous culture is often a primary metabolite, or the MO itself produced in the log phase, when the population is not under environmental stress

90
Q

in both batch and continuous culture?

A
  • as growth takes place, nutrients are used uo and waste products build up
  • as the culture reaches the SP, the overall growth ceases but end products still produced
91
Q

Bioreactor is a.k.a ?

A

fermenter

92
Q

B: why is there steam?

A

to clean valve

93
Q

B: nutrient or inoculant?

A

e.g. AA

94
Q

B: nutrient medium is ?

A

sterile, to get rid of comp from other species

95
Q

B: impeller?

A

mixes, preventing MO from settling at the bottom, motor causes it to turn

96
Q

B: O2 conc probe?

A
  • depends on MO and how they respire

* if are, high and if aner, low

97
Q

B: cooling jacket?

A

high level of metabolic activity and impeller moving which generates heat so needs to be cooled down

98
Q

B: cold water inlet and cold water outlet?

A

coldness maintained

99
Q

B: steam near harvest pipe?

A

steriliser of liquid being harvested

100
Q

B: antifoam?

A

preventing from foaming which could generate pressure

101
Q

B: pressure guage?

A

so doesn’t explode

102
Q

B: filtered waste gases?

A

to avoid removing MO too

103
Q

B: pH probe

A
  • pH change = enzymes denatured

* acid/ base added to alter pH

104
Q

B: T probe

A

T change can cause denaturation

105
Q

B: sparger?

A

air vent -allows bubbles ofgas to be put in. Can be used as a stirrer.

106
Q

B: compressed air?

A

the gas supply

107
Q

B: harvest pipe?

A

how products are harvested

108
Q

Advantages of batch culture?

A
  • Easy to set up - min attention needed during culture
  • fermenter can be used for diff cultures
  • only 1 batch needs to be discarded if cotamination happens
109
Q

disadvantages of batch culture?

A
  • down time between cultures

* large fermenters needed for profitable yields

110
Q

Advantages of continuous culture?

A
  • no down time

* small vessels can be used bc of continuous input and output

111
Q

disadvantages of batch culture?

A
  • sig problems if culture becomes contaminated

* culture can sometimes block input and output pipes

112
Q

What happens inside a bioreactor?

A

culture of MO grows

113
Q

what is a bioreactor?

A

vessel where conditions can be monitored and controlled very precisely

114
Q

hameocytometer?

A
  • used to estimate pop size of MOs
  • cells in a small precise V of suspension are counted
  • then answer multiplies up to the total volume of the pop
  • cover slip put over haemocytometer which sits exactly 0.1mm above the counting grid, so the liquid containing the cells to be counted is 0.1mm deep
115
Q

problems w culturing MO?

A
  • MOs potentially harmful - espec. if grown in anaer conditions
  • GM MOs must be contained inside bioreactors by law - biohazard centre
  • cultures easily contaminated with unwanted MOs
116
Q

What is done to avoid these issues?

A

aseptic techniques used

117
Q

e.g. of aseptic techniques - sterlilised?

A
  • all equip and agar sterilised beforehand using steam/ disinfectant/ ethanol
  • autoclave can be used for small items
118
Q

e.g. of aseptic techniques - protective clothing?

A

worn to prevent contamination FROM humans

119
Q

e.g. of aseptic techniques - methods which stop MO from settling down?

A
  • airflow upwards and out to prevent MOs settling on sterile surfaces
  • Lit Bunsen on bench
  • Higher pressure maintained inside bioreactor room with air pumps
  • Cultures prepared in fume cupboard
120
Q

e.g. of aseptic techniques - flaming?

A

• bottle openings and inoculating loops flamed

121
Q

e.g. of aseptic techniques - opening lids of agar plates and bottles?

A

kept to a min.

122
Q

e.g. of aseptic techniques - disposal of agar plates?

A

done carefully

123
Q

culturing MO- which conditions are prevented?

A

anaerobic, to prevent growth of dangerous toxin producing anaerobes

124
Q

other e.g. of aseptic techniques ?

A
  • steam cleaning

* washing w ethanol

125
Q

e.g. of enzymes used in industry?

A

baby food, biological washing powder

126
Q

immobilised enzymes?

A

can be done in a no. of diff ways so that the products can easily be separated again after the reaction, leaving the enzyme molecule to be reused

127
Q

IE - beads are?

A

totally porous - solution can go thru it

128
Q

Methods of immobilising enzymes?

A
  • adsorption to carrier
  • covalent boding to carrier
  • entrapment
  • encapsulation
129
Q

adsorption to carrier?

A

• enzyme bonded ionically or with hydrophobic interactions w carrier - gold, glass beads

130
Q

adv of adsorption to carrier?

A

+ simp and cheap
+ can be used w many diff processes
+ high reaction rates

131
Q

disadv of absorption to carrier?

A
  • bonds not that strong so some leakage can occur
132
Q

covalent bonding to carrier?

A

• enzyme molecules covalently bond to carrier and to other molecules

133
Q

adv of covalent bonding to carrier?

A

+ very little leakage because bonding is strong

+ enzymes very accessible to substrate

134
Q

disadv covalent bonding to carrier?

A
  • active site may be modified in the process, making it less efficient
  • cost varies
135
Q

entrapment?

A

• enzymes trapped in gel or network of cellulose fibres

136
Q

advantages of entrapment?

A

widely applicable to diff processes

137
Q

disadvantages of entrapment?

A

active site less easily available so reaction rates slower

  • can b difficult to entrap
  • effect of entrapment on enzyme activity very variable depending on matrix
138
Q

encapsulation?

A

• enzyme molecules separated from substrate by a partially permeable membrane, often in the form of a capsule

139
Q

disadvan of encapsulation?

A
  • may be expensive

- diffusion of the substrate to and product from the active site can be slow and hold up the reaction

140
Q

advan of encapsulation?

A
  • simple to do

- widely applicable to diff processes

141
Q

general advantages of IE?

A

+ Enzyme molecules stabilised by immobilisation - more resistant to high T and extreme pH

142
Q

general disadvantages of IE?

A

• Can slow reaction rates bc active site not as freely available as free enzyme

143
Q

Advantages of micropropagation?

A
  • sterile = infection would damage plant - disease free plant
  • large no. of gen identical plants produced
  • new plants which are seedless produced to customer’s tastes
  • new way of growing infertile plants
144
Q

disadvantages of micropropagation?

A
  • monoculture produced - plants are genetically identical - all susceptible to the same diseases
  • expensive, skilled workers req
  • explants and plantlets vulnerable to infection during production
  • if source infected by virus all clones will be infected

A Level Biology (28 decks)

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