Genetic Engineering

Question 1

Recombinant DNA

Answer 1

Combining 2 or more pieces of DNA not normally found together by artificial means

Question 2

Restriction Enzymes

Answer 2

1. Bacterial defense mechanism -> recognize short sequences of double-stranded DNA as targets
2. All have specific consensus sequence -> cleaving to a different base
3. 2 Different enzymes can give rise to compatible sticky ends

Question 3

Gel electrophoresis

Answer 3

1. DNA restrictive fragments placed ina well of an agarose gel -> charged
2. Molecules move through pores
3. Smaller travel further

Question 4

Southern blot

Answer 4

1. Target and probe combined by heat/ alkali then reannealed
2. Transfer to nitrocellulose
3. Hybridize with labeled DNA/RNA probe
4. This gives an audiodiagram

Question 5

Limitation of plasmids as vectors

Answer 5

Plasmids are limited in amount of DNA they carry -> yeast artificial chromosome 1000 kb

Question 6

Types of cell transfection

Answer 6

Transient and stable

Question 7

Difference between types of cell transfection

Answer 7

1. Transient -> cells only keep inserted genes for a couple of generations
2. Stable -> genes integrated into genome -> constant production of protein

Question 8

Xenotransplantation

Answer 8

Transfer of living cells from one organism to another

Question 9

CRISPRs

Answer 9

Clustered regularly interspaced short palindromic repeats

- Bacterial and Archean defence mechanism against plasmids and phages

Question 10

Cas9

Answer 10

CRISPR associated protein 9, a nuclease

Question 11

PAM sequence

Answer 11

DNA sequence immediately following the sequence targeted by Cas9

Question 12

Ethical considerations of GMO’s

Answer 12

1. Risk assessment and control measures
2. ST + LT impact
3. Balanced and informed decision making processes

Question 13

Western Blotting

Answer 13

1. Hybridize RNA polynucleotide
2. Mixture of proteins -> separate on gel and transfer onto membrane
3. Incubate protein sample with antibody 1 probe
4. Ligand-bind with antibody
5. If visualized then can view if antibody has bounded to antigen
6. Quantify protein -> monomeric, dimeric

Question 14

Proteomics

Answer 14

Study of protein population: analysis of genome compliments of proteins from DNA -> RNA -> protein