inserting target genes

Question 1

3 stages :

Answer 1

1. isolate target gene
2. insert target gene into vector
3. insert vector into bacteria

Question 2

describe how a gene is inserted into a plasmid

Answer 2

• the same restriction enzyme as was used to isolate target gene is used to cut plasmid
• this ensures sticky ends are complementary
• DNA ligase reforms phosphodiester bonds between sticky ends and plasmid
• forming recombinant DNA

Question 3

recombinant DNA

Answer 3

DNA from more than 1 source

Question 4

describe a vector is inserted into bacteria

Answer 4

• recombinant plasmid and bacteria put into ice cold calcium chloride
• heat shock
• increases permeability of bacterial cell wall

Question 5

when the target gene is isolated, what else should be added?

Answer 5

• promoter and terminator regions
• sticky ends
• marker gene