Biotechnology

Question 1

How is biotechnology implemented to our lives?

Answer 1

Food industry - Helps making lactose-free milk
Medicines - used to mass produce medicines such as penicillin and insulin
Agriculture - makes harvest more nutritious or have longer shelf life

Question 1

Cellular vs Molecular biotechnology

Answer 1

Cellular - Does not need deep understanding of DNA and RNA
Molecular - Needs to understand how to manipulate cultures in a DNA and sometimes RNA level

Question 2

What microbes are being used in biotechnology?

Answer 2

Virus, Archaea, Bacteria, Algae, Fungi

Question 3

What function does viruses play a part in biotechnology?

Answer 3

Bacteriophage - type of virus that infects bacteria by attaching to them and uses their vectors to carry genes to the host
T4 ligase - Source of enzyme to join two DNA strands

Question 4

Archaea in biotechnology

Answer 4

Used for their RNA polymerase as archaea lives in extreme environments meaning that their proteins are extremely thermostable, including the RNA polymerase. It can maintain activity in high temperatures

Question 5

Bacteria in biotechnology

Answer 5

Its main role are as host cells, meaning the cells that carries the new recombinant DNA. It is good at cloning (making identical DNA) and expressing proteins to make high value products or enzyme

Question 6

Algae in biotechnology

Answer 6

There is potential in using it as biofuels as it have a unique property of converting CO2 into hydrogen or ethanol, which are two components that are able to be used as fuels

Question 7

Fungi in biotechnology

Answer 7

The use of mold can be made into antibiotics, anticancer synthesis and immunomodulators. Yeasts can also act as a host to express proteins and grows fast

Question 8

E. coli vs Saccharomyces as host cells

Answer 8

E-coli - easy to tamper with the genes, can be a part of our normal flora, easy to extract genes, beneficial to digestion and immune regulation
Saccharomyces - better at expressing eukaryotic genes (since it is a eukaryote), generally recognised as safe (GRAS) which means that it can be digested

Question 9

Plasmid

Answer 9

Little circle of DNA typically found in bacteria. Can be easily gained or lost from the cell as it contains optional extra information

Question 10

Uses of plasmids

Answer 10

Common vector to introduce new DNA into a microbial cell

Question 11

Origins of plasmids

Answer 11

We can isolate plasmids from bacteria whether the bacteria is taken from soil, water or the human body. Plasmids can swap DNA easily with other bacterium (horizontal gene transfer), because of this bacteria can evolve.

Question 12

How to use plasmids

Answer 12

1. Replicating features - allows it to replicate independently
2. Selectable marker - lets plasmids be taken up by the bacterial cells by force
3. Cloning site - used to insert the foreign DNA because it is a site recognised by restrictions enzymes where it can cut up that part of the DNA to insert a new DNA in

Question 13

Cloning

Answer 13

Means to make identical organisms

Question 14

3 ways to clone

Answer 14

1. In vitro - using polymerase chain reaction (PCR) to replicate pieces of DNA
2. Propagation - look back on Cavendish bananas
3. Plasmid insertion - putting the DNA into a plasmid can make DNA be replicated

Question 15

Tools needed for DNA cloning

Answer 15

1. Copying DNA
2. Cutting DNA
3. Joining DNA

Question 16

Amplifying/Copying DNA enzyme

Answer 16

Using the process PCR where we can copy DNA using a thermostable polymerase from the archaea

Question 17

Cutting DNA enzyme

Answer 17

Restriction enzymes (endonucleases) are used to break down fragments of DNA (or mRNA). The same restriction enzymes needed to be used to cut the DNA and plasmids so that it recognises where the DNA should be at

Question 18

Joining DNA enzyme

Answer 18

To join the DNA fragment and the plasmid together, need to use ligase such as T4 ligase

Question 19

Steps to DNA cloning

Answer 19

1. Digestion and Ligation
2. Transformation and screening
3. Product: GMO

Question 20

First step to DNA cloning

Answer 20

Extract or amplify the DNA that we want (that produces the product that we want). Uses PCR to amplify/copy selected DNA. DNA is the ‘digested’ by cutting it into different pieces using a restriction enzyme and use the same restriction enzyme to cut the plasmid vector. Put the DNA fragment and plasmid together for ligation using the ligase to make a recombinant DNA

Question 21

What is in the ligation mixture?

Answer 21

Contains recombinant plasmids, non-recombinant plasmids and non-ligated DNA

Question 22

Why are there leftovers in the ligation mixture?

Answer 22

Because restriction enzymes and ligases are not 100% efficient so there will always be some starting plasma left over and bits of DNA that are not ligated

Question 23

Second step to DNA cloning

Answer 23

Transformation and screening is needed to get only the recombinant DNA out of the ligation mixture. Uptake of DNA (Transformation) be done through electric shock, heat or conjugation (transfers itself into a cell). Pours the mixture into a selective agar plate which enables the plasmid-containing cells to grow. Screens the cells of interest (the one with recombinant DNA) either phenotypically (fluorescent) or genotypically (using PCR for sequence based screen).

Question 24

Final product

Answer 24

GMO (genetically modified organism) is the final product. It is a recombinant gene carrying microbe.

Question 25

What are the risks of GMO?

Answer 25

It usually depends what foreign DNA is added into the organism. Some people may be allergic to the DNA that is inserted into the gene however overall it is very safe. However, dealing with dangerous genes, it needs to be handles in a lab or physical containment level. It also begs the question on legal constraints around GMO. Another risk if there is poor handling in these organisms and is able to make their way to our normal flora, it can be extremely harmful for the human body, animals or environment. Commercial risks: understanding whether the public can accept it as a product. So in short, there are ethical and public considerations to be made for GMO

Question 26

How is vaccine an example of a recombinant product?

Answer 26

Vaccines contain recombinant genes as the genes contained in vaccines are used to train the body’s immune system to recognise the target antigens. This type of vaccines contain genes of the target antigen

Question 27

4 types of vaccines

Answer 27

1. Live attenuated microbe - live microbe that will stimulate the immune system to respond
2. Kill microbes - Kill the cells, may still be quite antigenic, can also stimulate immune system
3. Antigen
4. mRNA coding for antigens - stimulating your body by presenting the antigen

Question 28

How does vaccine uses its genes to train the immune system?

Answer 28

It uses the antigen gene to express foreign protein (or antigen protein) in the body which is safe as this is only a surface protein of the virus (used to detect by immune system) that does not contain any other elements of the hepatitis virus