Lecture 16

Question 1

Method of manipulating DNA

Answer 1

• Identify gene/DNA of interest
• Isolate the gene into a vector
• Make the necessary changes to the DNA - join different DNA frags
• Re-introduce the construct into our organism of choice

Question 2

Restriction Endonucleases/Enzymes

Answer 2

naturally occurring enzymes,
some bacteriophage (virus) is unable to infect specific strains of E.Coli and is therefore restricted as the bacteria cuts it up

Question 3

Gel Electrophoresis

Answer 3

DNA has negatively charged back bone - will move towards the positive side, figure out the distance between bases

Question 4

DNA ligase

Answer 4

Joins molecules of DNA together, can join them in the lab. ends need to be complementary

Question 5

Plasmids

Answer 5

Circular DNA molecules, vectors, naturally occurring in bacteria, self-replicating (provide bacteria a selective advantage

Question 6

Why do we clone DNA?

Answer 6

Delete gene
Alter gene
Cross- species expression
Determine DNA sequence

Question 7

Generating constructs such as

Answer 7

• putting in or turning off a promotor
• find where a gene is
• create specific mutation

Question 8

Hybridisation

Answer 8

Denaturing DNA to single stranded DNA
Annealing (cooling) DNA, put them back together
label probe and make it bind