- The gene must be identified and isolated from Donor DNA (a gene that is isolated for insertion)
- The gene for insulin is isolated using a gene probe.
Using reverse transcriptase
- Functional mRNA coding for insulin is found within the pancreas.
- The mRNA is extracted.
- Reverse transcriptase (made by retroviruses) is added to make a complementary DNA copy
- The addition of DNA polymerase converts it into a double strand for insertion into a plasmid.
To insert a gene into a bacterium, a vector is used. Usually a plasmid. (circular DNA from a bacteria.
- The plasmid is obtained by dissolving the bacterial walls and plasmids are seperated from the debris.
- The plasmid is cut open using the same restriction endonuclease which hydrolyses the internal sugar phosphate bonds between nucleotides within the DNA. They…