P2 Cloning Organisms and Biotechnology Flashcards

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1
Q

How do plants clone themselves naturally?

A
  • By vegetative propagation: they reproduce naturally, without fertilisation, producing an offspring that is identical.
  • They do this using meristem (found in roots, shoots and between xylem and phloem).
  • A miniature plant (plantlet) is able to grow from these areas, once it grows enough to survive on it’s own, the plantlet detaches.
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2
Q

What are the methods of natural cloning in plants?

A
  1. Produce clones using horizontal stems, called runners (connecting the parent plant and plantlet).
  2. Produce clones using tubers (swollen, underground bits of stem). Buds form on the surface of tubers, which develop into clones (plantlets).
  3. Produce clones using bulbs (underground stems, enclosed by layers of leaves). Buds form between these layers in the bulb, which then develop into clones (plantlets).
  4. Produce clones using rhizomes (an underground, horizontal stem), which attaches the parent plant to the plantlet.
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3
Q

How can farmers encourage plants to naturally clone?

A

Using cutting:
1. Cut off bits of a healthy plants stem.
2. Dip the cut stem into growth hormones (eg. auxins).
3. Place the cutting in soil, and cover it with a plastic bag to create a moist, warm environment.

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4
Q

What are the advantages and disadvantages of using cuttings to grow plants?

A

Advantages:
1. No expensive
equipment
2. Doesn’t require much skill
3. Quicker than growing plants from seeds

Disadvantages:
1. All clones are identical, so they can all be killed off by a single disease, or change in the environment.
2. Only a limited number of clones can be produced from one plant.
3. Some plants don’t respond well to natural cloning.

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5
Q

How are plants artificially cloned?

A

Micropropagation:
1. Cut off a small sample of meristem tissue (containing totipotent cells) - this sample is called an explant.
2. Sterilise the sample to reduce the chances of contamination.
3. Place the sample in a sterile liquid containing growth hormones, which stimulate the cells to undergo mitosis. This creates a mass of cells called a callus.
4. The callus is then divided, and each part is placed into a new liquid containing hormones and nutrients.
5. These cells form plantlets, which are then potted.

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6
Q

Advantages and disadvantages of artificially cloning plants (micropropagation):

A

Advantages:
1. Rapidly produces a large number of clones.
2. Used to grow plants that are difficult to grow from seeds.
3. Plants can be grown at any times of the year.

Disadvantages:
1. All of the clones could be killed by a single disease/change in the environment.
2. Expensive process.
3. Requires skill to carry out.

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7
Q

How does cloning occur naturally in animals?

A

Identical twins:
- after fertilisation the embryo splits, forming twins

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8
Q

How can twins be created artificially?

A

Artificial embryo twinning:
1. Extract an egg cell from a female animal, and fertilise it in the lab.
2. As the embryo develops, it is split into single cells.
3. These single cells form new identical embryos, which are placed into different surrogate animals.
3. These surrogates give birth to genetically identical offspring.

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9
Q

How can adult animals be artificially cloned?

A

Using somatic cell nuclear transfer:
1. Extract a somatic body cell, then extract it’s nucleus.
2. From a different animal from that species, extract an unfertilised egg, and extract the nucleus from this egg (enucleation).
3. Insert the somatic cell nucleus into the enucleated egg, and apply an electric shock, which fuses the nucleus into the egg (electrofusion).
4. The transformed egg divides, and forms an embryo.
5. The embryo is transferred into the uterus of the surrogate animal, who gives birth.

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10
Q

Advantages and disadvantages of artificial cloning in animals (somatic cell nuclear transfer)

A

Advantages:
1. Can be used to clone animals with desirable characteristics eg. high milk yield.
2. Can be used to clone genetically modified animals that produce useful proteins for human medicine.
3. We can harvest stem cells from embryos grown in this way. Stem cells can then grow tissues and organs, which can be used for medical treatment.

Disadvantages:
1. Many cloned animals fail to develop.
2. Cloned animals often die early and have genetic abnormalities.
3. There are ethical issues with harvesting stem cells from embryos, as this destroys the embryo, which could have grown into an organism.

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11
Q

What is biotechnology?

A

The use of living organisms, such as microorganisms, to make products for humans.

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12
Q

What are the uses of microorganisms in biotechnological processes?

A
  1. Make food:
    - Bacteria is used to produce cheese and yogurt.
    - Yeast is used to make bread and ethanol.
  2. Make medicine, such as penicillin and insulin.
  3. Bioremediation: microorganisms break down contaminants in land and water, such as oils or pesticides.
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13
Q

Advantages and disadvantages of using microorganisms to make food

A

Advantages:
1. Microorganisms reproduce very quickly, so large numbers can be grown in a short amount of time.
2. Microorganisms can be grown on waste materials, reducing costs.
3. Microorganisms can produce healthier, high protein, low cholesterol products such as mycoprotein (a meat substitute).
4. Microorganisms can grow in low temperature environments, reducing costs.

Disadvantages:
1. Any contamination in the batch of microorganisms can cause food to spoil, resulting in health risks.
2. People may not want to eat food made using waste products.
3. Food produced has little flavour, so we need to add chemicals.

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14
Q

What is culturing, and what requirements does it need?

A

The process of growing microorganisms:
1. Nutrients (to provide the microorganisms with everything they need to grow and reproduce). This is provided from a liquid called a broth, or from a jelly-like substance called agar. This is called the nutrient medium.
2. Oxygen for aerobic respiration.
3. Optimum temperatures, to increase rate of enzyme-controlled reactions, without enzymes denaturing.
4. Optimum pH, to increase rate of enzyme-controlled reactions. Changes in pH cause the hydrogen and ionic bonds in enzymes to break.

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15
Q

What are the aseptic techniques used when culturing microorganisms?

A
  1. Wear gloves, to prevent microorganisms from our hands being transferred to the samples.
  2. Keep the agar covered as much as possible to prevent microorganisms from the air contamination the agar. Keep the plate sealed while the microorganisms are growing, and only open the plate for the minimum amount of time, holding the lid over the plate.
  3. Use fire to kill contaminating organisms, equipment is run through a flame before use, preventing microorganisms from the equipment entering the agar. And flame the mouth of the bottle once the sample is collected, killing any microorganisms that entered the bottle while the lid was off.
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16
Q

What is a dilution factor?

A
  • Total volume/volume transferred
  • It is a measure of how much of the original solution is in the next solution.

eg. 1cm3 transferred, added to 9cm3, dilution factor of 10.

  • Total dilution factor = all dilution factors multiplied together.
17
Q

How do you calculate the number of microorganisms in a culture broth when they cannot be counted due to being so many?

A
  • Carry out a serial dilution, take a sample of each solution and add it to a plate.
  • Allow the organisms to reproduce, then select the plate with the highest number of countable colonies.
  • Number of microorganisms per cm3 in the broth = number of colonies per cm3 x total dilution factor
18
Q

Formula to calculate the number of bacteria in a solution after divisions

A

N = N0 x 2^n

Where N = the number of bacteria after n divisions
N0 = initial number of bacteria
n = number of divisions

19
Q

What does a graph of the log of number of bacteria x time look like?

A

An exponential graph:
1. The lag phase is where the bacteria are taking in nutrients
2. The exponential phase is where the bacteria then divide exponentially
3. Stationary phase is when nutrients have run out so bacteria start dying, and the number of bacteria reproducing = the number of bacteria dying.
4. The death phase is then when the number of bacteria dying is greater than the number of bacteria reproducing.

20
Q

What are primary metabolites?

A
  • Substances that are produced by microorganisms that are directly involved in growth and reproduction.
  • Eg. ethanol is produced when yeast respires anaerobically.
  • These are mainly produced during the lag phase and exponential phase.
21
Q

What are secondary metabolites?

A
  • Substances that are produced by microorganisms that are not directly involved in growth and reproduction.
  • Eg. penicillin - secondary metabolites are typically produced after primary metabolites.
  • These are mainly produced during the stationary phase and death phase.
22
Q

How are microorganisms produced on a large scale?

A

Industrial fermentation:
- Fermenters are filled with a liquid containing nutrients.
- They have an inlet for air to allow oxygen to enter the tank.
- They have water running through a coat around the tank, keeping the tank at it’s optimum temperature.
- They contain a buffer to keep the pH constant.
- They contain a motorised stirrer, to keep the contents of the tank moving, helping the microorganisms to reach nutrients and oxygen.
- They contain gas outlets so that carbon dioxide produced during respiration can be released.

23
Q

What are the two methods of fermentation?

A
  1. Batch fermentation: microorganisms are added to the fermenter and left to reproduce until they reach the death phase. At this point the process is stopped and the products are harvested. The tank is cleaned, sterilised and a new nutrient medium and microorganisms are added - most appropriate for collecting secondary metabolites.
  2. Continuous fermentation: microorganisms are added to the fermenter and left to grow. When the microorganisms reach the exponential phase they are removed from the tank and immediately replaced with new microorganisms and nutrient medium - most appropriate for collecting primary metabolites.
24
Q

What are immobilised enzymes?

A

Enzymes bound to an insoluble material, making it easier to collect the enzyme at the end of the reaction.

25
Q

What are the 4 methods of immobilising enzymes?

A
  1. Adsorption:
    - enzymes are bonded to a surface, such as clay, through ionic bonds and hydrophobic interactions.
  2. Covalent bonding:
    - enzymes are bonded to a surface, such as clay, through covalent bonding.
  3. Entrapment in a matrix:
    - the matrix is a material eg. collagen, that prevents the enzyme from moving
  4. Membrane separation:
    - the enzyme can be separated from the reaction mixture with a partially permeable membrane
    - the substrate diffuses through the membrane to reach the enzyme, and the product diffuses out
26
Q

Advantages and disadvantages of using immobilised enzymes

A

Advantages:
1. Reduces downstream processing (easier to separate the enzyme from the product).
2. Allows the enzyme to be recovered and reused, reducing costs.
3. Immobilising enzymes increases their temperature tolerance, so the reaction vessel can be heated to higher temperatures, increasing rate of reaction, and therefore yield.

Disadvantages:
1. Expensive to immobilise enzymes.
2. Enzymes may detach and leak into the reaction mixture.
3. Immobilisation may change the shape of the enzymes active site, reducing rate of reaction.

27
Q

Uses of immobilised enzymes

A

Used in biotechnology