Production of Insulin

• Step 1 (Production of Insulin)

  • Human insulin gene is isolated
  • Insulin gene is isolated so that segment of gene that is inserted is very small
  • Restriction enzyme cuts the restriction site of the gene
  • Sticky ends are produced at both ends of the gene

  
• Step 2 (Production of Insulin)

  • A plasmid is removed from a bacterium (E.coli)
  • Plasmid is cut with the same restriction enzyme
  • Same restriction enzyme is used so that same sticky ends at human insulin gene and plasmid have complementary shape
  • The sticky ends produced complementary to those of the insulin gene

  
• Step 3 (Production of Insulin)

  • Insulin gene and the cut plasmid bind by complementary base pairing between their sticky ends
  • DNA ligase binds adjacent nucleotides and seals the gene to the plasmid
  • A recombinant plasmid is formed

  
• Step 4 (Production of Insulin)

  • Recombinant plasmid enters an E.coli bacterium
  • A transgenic bacterium is produced
  • E.coli bacteria is used because bacteria are single celled and rapidly reproduce, increases insulin production

  
• Step 5 (Production of Insulin)

  
  

  • Since bacteria should not be injected into insulin patients, insulin hormone is extracted by lysis
  • Then it is purified to remove bacteria cell protoplasm
  • Lysis - breakdown of cell cause by damage to its plasma membrane
  • Bacteria is cultured using a bioreactor so that more insulin manufacturing bacteria can be produced