3. In Vivo cloning

Question 1

define DNA ligase

Answer 1

enzyme which catalyses the joining of phosphate groups and sugars together

Question 2

define recombitant DNA

Answer 2

foreign DNA which has joined with a vector

Question 3

define a vector in terms of gene technology

Answer 3

anything which carries recombinant DNA into the host cell

Question 4

what does in vivo mean

Answer 4

anything that takes place within an organism

Question 5

outline the features of in vivo cloning

Answer 5

• inside an organism
• DNA fragments placed into plasmid
• takes longer
• not all plasmids take up the gene
• requires marker genes to see if DNA fragments have been taken up

Question 6

define a promoter

Answer 6

something added to the gene which signals RNA polymerase to start

Question 7

define a terminator

Answer 7

signals RNA polymerase to stop

Question 8

outline the process of in vivo cloning

Answer 8

1. restriction endonucleases isolate the target gene
2. promoter and terminator genes are added onto the target gene
3. restriction endonucleases cut specific sticky ends at complimentary recognition sites, allowing them to join
4. ligase enzymes catalyse phosphate and sugars joining together, fusing the DNA fragment and plasmid
5. not all plasmids insert the DNA fragments
6. the host cells multiply, if they took up the gene, they will have a marker gene present

Question 9

what are the ethical issues of in vivo cloning?

Answer 9

• may cause harmful mutations
• may question whether altering DNA can be done to humans