gene technology

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describe the steps involved in the production of bacteria capable of synthesizing human insulin.

1) identifying the human insulin gene

  • beta cells in islet of langerhans in the pancreas produced large amounts of insulin and will therefore have large amounts of mRNA for it
  • mRNA has a genetic code complementary to the exons (key portions of DNA - which are translated into proteins - useful part) of a human insulin gene
  • mRNA contains more exons than DNA therefore works faster

2) isolating mRNA and making CDNA using reverse transcriptase

  • mRNA is isolated from beta cells
  • mRNA is incubated with a mixture of free DNA nucleotides and reverse transcriptase ( from viruses - use RNA as their genetic material)
  • produces single strand of DNA - CDNA - complementary DNA

3) cloning the DNA using DNA polymerase

  • single strand is made double by DNA polymerase
  • cloned to make many CDA molecules using polymerase chain reaction (PCR)
  • PCR makes DNA amplification possible
  • heat => 'melting' - hydrogen bond breaks
  • cool - hydrogen bonds begin…


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