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Biology > 6.4 - Cloning and biotechnology > Flashcards

6.4 - Cloning and biotechnology Flashcards

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1
Q

What is a clone?

A

A genetically identical organism or cell

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2
Q

How do plants clone themselves naturally?

A

Vegetative propagation

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3
Q

What is vegetative propagation?

A

When plants send out a branch (Normally underground, through the roots) in order to create another plant

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4
Q

What are the different ways that plants can perform vegetative propagation?

A
  • Roots can branch off
  • Runners can branch off (Slightly above ground but similar to roots)
  • Rhizomes (Similar to roots)
  • Stolons (Similar to roots)
  • Suckers (Similar to roots)
  • Bulbs (Via overwintering organs: Onions, potatoes etc)
  • Corns (Via overwintering organs: Onions, potatoes etc)
  • Tubers (Via overwintering organs: Onions, potatoes etc)
  • Via the leaves dropping off to form a new plant
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5
Q

Why do plants perform vegetative propagation?

A
  • Vegetative propagation is fast as a partner isn’t needed
  • There is no genetic variation, If the plant has desirable alleles then this is a benefit as the desirable characteristics will be passed on
  • However this could mean that the plants are all susceptible to the same diseases or they could all suffer from climate change as there is no genetic variation and evolution cannot occur within clones to prevent extinction/borderline extinction
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6
Q

Why is the process of artificially cloning plants so easy?

A

The process is easy as totipotent meristem cells are in many different parts of the plant

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7
Q

What does totipotent mean?

A

-When a cell is mostly undifferentiated and it has the ability to divide into all the different cell types it is described as totipotent
(When a cell is the most undifferentiated)

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8
Q

What are the different ways of artificially cloning plants?

A
  • Cutting, this is when a cutting of a plant is taken and planted in soil to grow
  • Micro propagation/Tissue culture, growing large numbers of new plants from meristem tissue taken from a sample plant
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9
Q

What is the process of artificially cloning plants via cutting?

A
  • Cut plant between the nodes using a sharp blade (At 45 degrees)
  • Dip the cutting in a powder of a rooting hormone (Normally auxin)
  • Pot the cutting in moist soil
  • Cover the leaves in a growing box or a plastic bag to increase the humidity of the air
  • This reduces transpiration stress
  • Leave the roots to develop
  • The plants can then be re-potted
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10
Q

What is the process of Micro propagation/Tissue culture?

A

1) Use a sterile scalpel to remove the growth tip of a plant (Containing meristem cells)
2) Transfer the growth tip to the sterile nutrient agar
-To speed up the cloning and to make it more effective:
[Take the callus that is formed on the agar and cut it up into smaller pieces]
{The small pieces of the callus will each develop into different plants}
3) Apply shoot stimulating hormones to the growth tip
4) Apply root stimulating hormones to the growth tip
5) This forms tiny plantlets from the growth tip which are clones of the sample plant

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11
Q

Why must Micro propagation/Tissue culture be done aseptically?

A

As agar jelly is used the process must ensure there is no contamination

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12
Q

Why must the agar used in Micro propagation/Tissue culture contain nutrients for the growth tip?

A

The nutrients are used by the growth tip for respiration as the growth tip will not be able to produce sufficient nutrients from photosynthesis

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13
Q

What are the advantages of artificial plant cloning?

A
  • The process is rapid
  • Can reproduce plants that are sterile/seedless (Bananas and orchids are micropropagated to reproduce large amounts of sterile offspring)
  • Plants are genetically identical, this is an advantage as it is easy to know what each plant needs to survive and be healthy
  • Also, as the plants are genetically identical it means that the phenotype is predictable, this can be helpful when producing crops that are sold as the growth and characteristics of the plants are predictable and all the same
  • Another advantage of artificial cloning is that diseased plants can be cloned without passing the disease on to the offspring
  • This can be done by using meristem tissue from a different part of the plant that is disease-free and can be used to create a disease-free clone
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14
Q

What are the disadvantages of artificial plant cloning?

A

-It takes skilled labour to perform artificial plant cloning
(Labour intensive)
-It is expensive (Due to equipment)
-It must be done under aseptic conditions to prevent contamination and mould from growing on the agar
-There is a disease susceptibility in the offspring as there is a lack of genetic variation

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15
Q

How are animals naturally cloned?

A
  • Identical twins are natural animal clones
  • Identical twins are made when a single egg (zygote) is fertilised, the egg then divides in 2 to create identical twins who share the same genes
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16
Q

Why are animals artificial cloned?

A
  • Reproductive cloning is used to create clones of an animal which has desirable characteristics
  • The desirable characteristics can be passed down through alleles to the clone
  • Artificial cloning of animals can also be used to clone endangered species in order to increase the population size
  • Cloning can be done for research
  • Cloning can be done to reproduce offspring of genetically engineered animals that can be used to produce pharmaceuticals
  • Lastly artificial cloning can be used for therapeutic cloning
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17
Q

What are the different ways of reproductive cloning?

A
  • Embryo splitting

- Somatic cell nuclear transfer

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18
Q

What is the process of embryo splitting?

A

[E.g: In cows]
-An egg is taken from a cow with desirable characteristics and sperm is taken from a bull with desirable characteristics
-The egg and sperm go through IVF
-The embryo that is made from the IVF is split up using a scalpel blade controlled with dials (Divided)
-The embryos are then inserted into surrogate mothers’ uterus
-The baby calves are then born identical with the same genetic make up
(They are clones of each other, not clones of the parents)
-This process is done to make many offspring with the genetics of two parents which have desirable characteristics

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19
Q

What is the process of somatic cell nuclear transfer?

A

-Skin cells (2n diploid) are taken from the animal that is being cloned
-An egg cell (n haploid) from the egg donor is taken and enucleated
[This removes the nucleus, and the genetic material from the egg]
-The nucleus from the skin cell is fused with the enucleated donor egg cell
[This is done via an electric shock, the electricity allows the nucleus to be transferred into the egg cell as the egg cell is triggered to think it has been fertilised]
-The zygote (fertilised egg cell) is inserted into the temporary oviduct while it is forming into the embryo
-The embryo is formed from the zygote multiplying by mitosis
-The embryo is then inserted into a surrogate mother which then births the clone
(The offspring has the exact same DNA as the animal that gave the skin cells at the start of the process.

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20
Q

Why is the DNA of the offspring not 100% genetically identical to the animal that was to be cloned at the start of the process of somatic cell nuclear transfer?

A
  • There is mitochondrial DNA that remains from the egg cell donor even after it is enucleated
  • This is because mitochondrial DNA is always passed down from the egg cell in the mother
  • This contributes to a very small fraction of the offspring’s genetic information
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21
Q

How can somatic cell nuclear transfer be used for therapeutic cloning?

A
  • When someone is injured and damages cells in their body that are needed they could use somatic cell nuclear transfer to clone their cells and recover from the injury
  • A skin cell would be taken and cloned
  • The cell would, however be stopped before it develops into an embryo as it would just be cloned to take stem cells for therapies
  • The stem cells will be injected into the injury so they can differentiate and help the person recover from the injury with the development of the new cells
  • There is no chance of rejection as the stem cells are genetically identical to the person who is having them for therapy
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22
Q

How can cloning be used for research and drug testing?

A
  • The clones will all have identical DNA, this means that there is a controlled variable which cannot effect the outcome of research
  • This is used when testing a drug on a large number of animals that are all clones of each other to see if the drug is viable
  • Cloning is also used for drug testing as clones of animals with certain conditions would be made, the clones are then all tested with different drugs to see the effectiveness of the drugs in treating the condition
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23
Q

What are the advantages of animal cloning?

A
  • Whole herds of animals with ideal characteristics can be cloned to have genetically identical DNA
  • Cloning is used for research as the effect of genetics is controlled
  • Medical testing is done on clones with identical DNA
  • Endangered animals can be cloned
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24
Q

What are the disadvantages of animal cloning?

A
  • The clones have a lack of variation meaning that diseases could effect them all and potentially cause extinction
  • The success rate is low, this means it is unethical for animals and would be unethical to do it with humans
  • There are ethical issues with therapeutic cloning and killing embryos or stopping the development of embryos in somatic cell nuclear transfer when making stem cells for therapy
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25
Q

What is biotechnology?

A

Using living organisms or parts of living organisms in an industrial process

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26
Q

What is an example of a part of a living organism that is used in industrial processes?

A

Enzymes are parts of living organisms that are used for industrial processes

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27
Q

What are the advantages of using living organisms in industrial processes?

A
  • Microorganisms are cheap and easy to grow
  • Microorganisms can survive in low temperature and pressure
  • Fermenters can be built anywhere (Irrespective of the outside climate)
  • Microbes can be fed agricultural waste
  • Microorganisms reproduce quickly
  • There are no real ethical concerns with using microorganisms
  • Products are easy to isolate/retrieve from the culture medium
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28
Q

What are the disadvantages of using living organisms in industrial processes?

A
  • There is a risk of contamination when using microbes as, if the wrong bacteria (Contaminated bacteria) enters it will reproduce and cause a contaminated culture which can produce the wrong product and be harmful (Especially when producing food/drink)
  • It can be hard to isolate the protein
  • SCP has a high purine content which can cause gout, this means it must be reduced
  • Additives are required to make the product taste better
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29
Q

How is bread made using biotechnology?

A

-Bread is made from yeast with the microbe saccharomyces cerevisiae
-The yeast anaerobically respires the glucose from dough to produce ethanol + CO2
Glucose -> Ethanol + CO2] {The ethanol evaporates off}

30
Q

How is cheese made using biotechnology?

A

-Cheese is made from a bacteria called lactobacillus
-The lactobacillus respires to break down the sugars in milk
[Lactose -> Lactic acid] {This lowers the PH}
-The enzyme “Rennin” separates the protein from the milk
-The milk coagulates into curds and whey
-The curds are used to make up the cheese

31
Q

How is beer made using biotechnology?

A
  • Beer is made from yeast with the microbe saccharomyces cerevisiae
  • Barley grains are partially germinated and boiled down
  • This creates a sugary liquid containing glucose
  • Yeast anaerobically respires the glucose into ethanol + CO2
  • [Glucose -> Ethanol + CO2]
32
Q

How is yoghurt made using biotechnology?

A
  • Yoghurt is made from a bacteria called lactobacillus
  • Lactobacillus acidifies the milk, causing the PH to drop and proteins to denature
  • This changes the texture of the milk into a thicker “yoghurt” texture
  • Probiotic yoghurt drinks have other bacteria added which could be beneficial for the intestine
33
Q

Where does the enzyme “Rennin” come from?

A
  • Typically rennin comes form calves stomachs
  • However now some rennin is genetically engineered so that the cheese is vegetarian as the rennin does not have to be taken from a calves stomach
34
Q

What is single cell protein?

A

Single cell proteins are proteins derived from a culture of single-celled organisms, used especially as a food supplement

35
Q

How are single cell proteins used to make food?

A

-A single celled organism such as a fungus is grown in a tank
-The tank must have a maintained climate and the fungus must be supplied with nutrients into the tank
-As the fungus grows, the biomass of the single celled organism increases
-Once the SCP is finished growing, the tank is drained and the cells are collected and moulded into food
(E.g: Quorn)
-The purine content of the SCP may have to be reduced before it is safe to eat large quantities of it
-Additives might need to be added
-This process can be done anywhere with a tank to contain the SCP as it grows

36
Q

What is “air protein” and how is it made?

A
  • Air protein is a bacteria that is fed H2 and CO2 gas in order for it to grow
  • The H2 is produced from electrolysis of water (Splitting of the water molecule, artificially)
  • The CO2 comes from the air
  • The bacteria grows and produces a protein rich flour which can be added into foods for humans or it can be used to feed animals in agriculture
37
Q

Why does wine have a 12% alcohol content?

A
  • At roughly 12% alcohol yeast dies

- Distillation is used to produce more alcoholic drinks

38
Q

What would an industrial fermenter be used for?

A
  • An industrial fermenter is used for:
  • The brewing of beer/wine
  • Growing single cell proteins
  • The production of drugs like antibiotics (E.g: Penicillin)
  • The production of insulin (Using GM bacteria)
  • The production of enzymes that are extracted from living organisms
  • The production of citric acid (A food additive)
  • Biogas (The renewable energy source)
39
Q

What is an industrial fermenter?

A

An industrial fermenter is a high control growth vessel for a microbe

40
Q

What are the parts of an industrial fermenter and what are the purposes of each part?

A

-Mixing blades, move the microbes around so they have roughly the same exposure to O2 and nutrients and to prevent them from settling to the bottom
(At the bottom they would have to compete with each other for nutrients and O2 etc)
-Nutrient medium (Inlet), to feed the nutrients into the fermenter. The inlet must be sterile to prevent the entering of contaminating microbes which could cause harmful disease or cause growth of the wrong microbe
-Air inlet, where the air enters the fermenter. The air inlets must also be sterile to prevent contamination
-Sparger, this is to bubble the air in the fermenter to mix it into the culture
-Water jacket (Water in/out around the fermenter), this is to maintain the optimum temperature for the microbe in the tank (The water does not actually mix with the culture)
-Sensors, to detect and monitor: PH, temperature and minerals
-Pressure outlet valve, to release the gases out the tank and reduce the pressure inside the tank
-Outlet to drain tank, this could be to remove the product at the end

41
Q

What is a continuous culture in an industrial fermenter?

A
  • The nutrients in are the same as the product out constantly
  • This creates a constant rate of microbial growth
  • This is beneficial for primary metabolites
42
Q

What is a batch culture in an industrial fermenter?

A
  • The whole tank must be drained once the product has finished growing and is ready
  • The industrial fermenter must then be set up again for the next cycle of fermentation (Growth of a microbe)
  • Batch culture is used to get bacteria into the secondary phase to produce secondary metabolites
43
Q

What are the phases that a bacteria goes through when it is placed in a new growth medium?

A
  • The lag phase, the bacteria are acclimatising to the new climate. This could mean that genes are having to be turned on/off to acclimatise to a new environment (There are low numbers of bacteria)
  • The log phase, this is where the population is doubling as the bacteria reproduces (Doubling as quick as every 20 minutes)
  • Stationary phase, when there is a factor that limits the growth (E.g: O2 or nutrients)
  • Death/decline phase, the population is decreasing as bacteria are dying. This is caused by a factor that is severely lacking such as nutrients
44
Q

What is the formula that is used to work out the size of a population after a certain amount of time?

A

N = No x 2^n

N0 = Starting size of the population
n = How many doubles
45
Q

There was 100 bacteria at the start of the growth
The bacteria was left for 12 hours
The bacteria could double every 20 minutes

How many bacteria were there at the end of the 12 hour period?

A
  • The bacteria could divide 3 times an hour
  • 3 x 12 = 36 times in 12 hours
N = 100 x 2^36
N = 6.87 x 10^12
46
Q

How is a batch culture carried out?

A
  • The tank/fermenter must be sealed
  • Wait for growth
  • The tank must be drained to retrieve the finished product
47
Q

What is secondary metabolite?

A

A product that is only produced in stationary or decline phase (E.g penicillin)

48
Q

Why is penicillin a secondary metabolite?

A
  • Penicillin is a secondary metabolite as it is only produced in the stationary or the decline phase
  • This is because penicillin is only produced to kill other microbes when the fungus has high competition
49
Q

Why is a batch culture needed to produce secondary metabolites?

A
  • Because the culture must be in the stationary or decline phase to produce secondary metabolites
  • Continuous culture would not be able to produce secondary metabolites because the culture is maintained in the exponential phase as nutrients are being pumped in at the same rates that products are being removed, so products are removed after the exponential phase
50
Q

What are the advantages of batch culture?

A
  • It is the only way to produce secondary metabolites
  • There is less risk of contaminating produce, this is because the produce is being removed after the growth is finished. This means that the tank is sterilised after each batch so if contamination occurs it would only affect one batch as opposed to the whole on going produce (All on going produce would be contaminated if there was contamination in continuous culture)
51
Q

What is a disadvantage of batch culture?

A

Time is wasted when the culture is removed and the tank is sterilised after each batch, this makes the whole process of producing the cultures less efficient

52
Q

What are the main, major features of aseptic technique?

A
  • Doors and windows must be shut, this is to reduce the movement of bacteria in the air which would cause contamination
  • All surfaces, tools, clothes, hands etc must be sterilised/washed and clean
  • Hair must be tied back
  • A fume cupboard could be used to reduce the risk of contamination, this is because the fume cupboard would produce a constant flow of filtered air which prevents particles of pathogens from landing/settling on the culture
  • Work quickly and efficiently
  • Don’t put equipment down on surfaces
  • Use a flame to sterilise/re-sterilise
  • Lids should be put on/off quickly at an angle to cover the petri dishes or bottles. This is done to reduce the chance of pathogens entering the culture and causing contamination
53
Q

What is an error that can be done when transferring a culture using aseptic technique?

A
  • You must wait for the inoculating loop to cool down after it has been flamed to sterilise it
  • This is because if it is too hot it can kill the bacteria that it is dipped in
54
Q

What is the dilution practical used to work out?

A

The dilution practical is used to determine the number of viable bacteria in a culture

55
Q

How would a serial dilution be used to dilute a sample of Yakult in order to get a smaller sample of bacteria?

A

1) The starter culture will be Yakult
[Just Yakult in test tube 1]
2) The starter culture is diluted x10^-1
[9ml of sterile broth and 1ml Yakult in test tube 2]
3) The next is diluted x10^-2
[9ml of sterile broth and 1ml from test tube 2 is in test tube 3]
4) The next is diluted x10^-3
[9ml of sterile broth and 1ml from test tube 3 is in test tube 4]
5) The next is diluted x10^-4
[9ml of sterile broth and 1ml from test tube 4 is in test tube 5]
-Etc…

56
Q

When researching for the number of viable bacteria in a culture, what is done after the serial dilution?

A

-Take a sample (100 micro litres) from each test tube and put it onto an Agar plate
[1ml = 1000 micro litres]
-Place the cultures in an incubator at 32°C and wait 24 hours
-After the 24 hours, take the cultures out and observe the plates
-Count the colonies of bacteria on the agar plates that have visible colonies without a lawn of bacteria
-If 18 colonies were counted, then it means that within the 100 micro litre solution, there were 18 bacteria that landed on the plate that were viable and started to grow

57
Q

Why are the cultures left inside of the incubators at 32°C

at a school rather than 37°C?

A
  • The culture is likely to grow pathogens at 37°C so they must be kept at a lower temperature at schools as it is safer
  • This is because the culture has less chance of growing pathogens at the lower temperature of 32°C
58
Q

What is the formula to work out the number of bacteria in the original Yakult sample?

A

18 in 100 micro litres of the x10^-4 dilution
18 in 0.1 ml so…
180 bacteria in 1ml
[As this is the 10^-4 test tube it is 10000 times more diluted than the Yakult]
180 x 10000 = 1.8 million colony forming units per ml in the original broth

59
Q

What should be done if the most diluted culture is still showing a lawn of bacteria?

A

If the most diluted agar plate shows a lawn of bacteria then it is necessary to do another dilution series to get more diluted samples

60
Q

Why is it better to take a reading of the number of colonies from a less diluted culture? (Presuming it isn’t showing a lawn of bacteria)

A
  • Taking a reading from a culture with more colonies could give a more precise estimation of the number of viable bacteria in the original culture
  • Taking a reading from a petri dish with 18 colonies is gonna be less precise than taking a reading from a less diluted sample which shows 183 colonies
  • This is because the petri dish showing 18 colonies would predict that there are 1.8 million CFU per ml but the less diluted sample of 183 would predict that the original culture would have 1.83 million CFU per ml which is a more precise estimation
  • Also the more diluted series’ are more error prone due to rounding which would need to be done for the estimation of the original culture
  • This is shown as a more diluted series would have 1.8 colonies which could be rounded to 2, this shows that more diluted series’ are less precise due to rounding up/down which can affect the accuracy/preciseness of the estimation for the original culture
61
Q

Why is it advised to not take a sample from a very undiluted culture when working out an estimation of the number of colonies from an original culture?

A
  • It can take a long time to count the number of colonies as they will be higher in a less diluted culture
  • It can also be harder to distinguish between the different colonies as a lawn of bacteria can be seen to cover the agar plate
62
Q

What are the advantages of using immobilised enzymes for biotechnology?

A
  • Enzymes can survive and grow at low temperatures and pressures so it is easier to maintain and grow them
  • Immobilised enzymes don’t mix with the other products that are made as they are fixed and cannot move, this means it is easier to separate the product from the enzyme
  • Immobilised enzymes make the continuous process easier, this is because a liquid containing substrates can flow over a surface with immobilised enzymes causing the substrates to bind to the enzymes and the product to flow out the fermenter continuously
  • As the enzymes are fixed and cannot move, it is easier to reuse the enzymes
  • When enzymes are immobilized and attached to a surface it makes them more resistant to temperature and PH changes (They are more resistant to denaturing)
63
Q

What are immobilised enzymes?

A

Enzymes that are fixed to an inert, insoluble material so that they cannot move

64
Q

What are the different ways of immobilising an enzyme?

A
  • Adsorption
  • Covalent bonding
  • Entrapment
  • Membrane separation
65
Q

What is adsorption?

A
  • Adsorption is sticking an enzyme to a clay particle to cause a temporary bond (E.g: Using hydrophobic interactions and ionic or hydrogen bonds as they are more temporary)
  • The enzyme can potentially become unstuck from the clay particle, this can cause the enzyme to leak into the mixture
  • Adsorption is easy to carry out and immobilise the enzyme, however it is semi-permanent as the enzyme can become unstuck and leak into the mixture
66
Q

How are enzymes immobilised from covalent bonding?

A
  • Covalent bonding is a more permanent and more expensive way of immobilising enzymes
  • An insoluble, inert material is covalently bonded to the enzyme in order to stop it from moving (Immobilise it)
  • Covalent bonds can sometimes bond to the enzyme over the active site, this blocks any substrates from bonding to the enzyme so it reduces the activity of it
67
Q

What is entrapment?

A

-Entrapment is when the enzymes are immobilised in a gel/matrix
-The network of gel holds the enzymes in place as they cannot move and are immobilised
(E.g: Enzymes can be tangled in a cellulose mesh which entraps the enzymes in order to immobilise them) [Prevent them from moving]

68
Q

What is membrane separation?

A
  • Membrane separation is when enzymes are blocked from exiting as they are too big to travel through the membrane
  • This can happen when enzymes bond with a substrate to make a product, the product is able to leave the membrane as it is small enough however the enzyme cannot as it is too big so it is immobilised and prevented from leaving the membrane
69
Q

What are some examples of immobilised enzymes?

A
  • Glucose isomerase is an enzyme that converts glucose into fructose
  • Glucose isomerase is useful for making dieting foods as fructose is a sweeter sugar so less is needed meaning that the food is healthier
  • Lactase is an enzyme that converts lactose into glucose and galactose
  • Lactase is useful for people who are unable to digest lactose, lactase can be used to make lactose free milk. The immobilised enzyme, lactase works to break down the lactose into glucose and galactose so it can be digested by someone who is lactose-intolerant
  • Aminoacyclase is an enzyme that produces amino acids for chemical synthesis
  • Glucoamylase is an enzyme that converts starch into glucose, this can be used with foods or drinks that are high in starch contents
  • Corn syrup that is high in starch is treated with glucoamylase in order to produce glucose, this decreases the starch contents as the starch is converted to glucose
  • Nitrile hydratase is an enzyme that converts nitriles to amides
  • Nitrile hydratase is used to produce monomers that can be used in polymerisation to make the polymer “Polyacrylamide”
70
Q

What are the practical techniques that are on the specification for “6.4 Cloning and biotechnology”?

A
  • Making a cutting
  • Performing tissue micropropagation
  • Using aseptic technique
  • Making a serial dilution (Dilution plating experiment)
  • Practical techniques used for cloning
71
Q

What is micropropagation/tissue culture?

A
  • Growing large numbers of new plants from meristem tissue taken from a sample plant
  • Growing new tissues organs or plants from certain tissues cut from a sample plant

Biology (23 decks)

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