Genetic engineering

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  • 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

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