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A-Level Biology OCR A

Insulin Gene Plasmids Resistance Bacteria Human Dna Ends

This flashcard outlines the process of genetic engineering to produce human insulin using bacteria. It details the steps involved, from mRNA extraction to culturing the modified bacteria and extracting the insulin.

Cette fiche décrit le processus de génie génétique pour produire de l'insuline humaine à l'aide de bactéries. Elle détaille les étapes, depuis l'extraction de l'ARNm jusqu'à la culture des bactéries modifiées et l'extraction de l'insuline.

Front Describe the process of genetic engineering to produce human insulin using bacteria.
Back Producing human insulin:
  • mRNA extracted from beta cells and used to make double stranded DNA
  • Nucleotides added to the ends to form sticky ends
  • Vector used is a plasmid already modified to contain amipicilin & tetracyline resistance genes
  • Plasmids are cut by restriction endonuclease in middle of tetracycline resistance gene & insulin gene is inserted using DNA ligase
  • Plasmids are inserted into bacteria through heat shock/electroporation
  • There are three possible types of bacteria formed:
    • Took up no plasmids - no resistance
    • Took up plasmids without insulin gene - resistance to both antibiotics
    • Took up plasmids with insulin gene - only resistant to ampicilin
  • To extract insulin gene containing bacteria, serial dilute bacteria and transfer to agar to grow colonies from single individuals, test resitance of some individuals from each colony to find those with insulin gene
  • Place successful colonies in incubator with optimum conditions, extract insulin

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