BIOC192 Lecture 19 - Recombinant DNA technologies

Question 1

what are recombinant DNA technologies?

Answer 1

joining bits of DNA together (sometimes from different species). These are then inserted into an organism to produce (express) a useful protein

Question 2

what are the 3 key components of recombinant DNA plasmids?

Answer 2

1) origin of replication (ORI)
- allows initiation of replication using host DNA polymerase
2) antibiotic resistance gene
- allows selection of cells containing plasmid
3) promoter
- drives expression of your favourite gene (e.g insulin or GFP) in cells with appropriate transcription factor machinery

Question 3

what was the insulin treatment method in the 1960’s?

Answer 3

• insulin first isolated from pig and cattle
• insulin protein sequence/structure (not identical)
• not always 100% pure

Question 4

how do you cut and pst DNA into plasmids?

Answer 4

• start of with a naturally occurring bacteria plasmid
• then insert the promotor in our ‘favourite’ transgender of the bacterial plasmid to be used for recombinant DNA technologies