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Production of Human Proteins

Production of Human Proteins. Avery Grant and Eliza Tarwater. Diabetes- Type 1 Diabetes Mellitus. Metabolic disease Cause: lack of insulin Treatment: insulin injection. Treatment. Older. Newer. Pancreatic tissue from pigs and cattle Expensive Side effects Often contaminated.

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Production of Human Proteins

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  1. Production of Human Proteins Avery Grant and Eliza Tarwater

  2. Diabetes- Type 1 Diabetes Mellitus • Metabolic disease • Cause: lack of insulin • Treatment: insulin injection

  3. Treatment Older Newer • Pancreatic tissue from pigs and cattle • Expensive • Side effects • Often contaminated • GM Human Insulin • Genetically modified bacteria and yeast • Since 1980s • Greatly improved management of type 1 Diabetes

  4. Synthesis of Human Insulin Using Recombinant DNA Technology • All involve • putting a human gene into a plasmid (which can either be a yeast or bacteria) • secreting a protein which insulin can be derived from

  5. STEP 1 • Identify and Synthesize a human gene • Insulin: • Small protein • 51 amino acids- A&B • polypeptide chains • STEP: Chemically synthesize DNA chains that have nucleotide sequences for A and B “genes”

  6. STEP 2 • Insert the Synthetic DNA into Plasmids • A and B nucleotide sequences individually put into gene for bacterial enzyme • Makes bacteria produce insulin

  7. STEP 3 • Put Plasmid in Bacterial Cell • Recombinant plasmids introduced to E. coli • Requires millions of copies of bacteria w/ plasmid of insulin gene • Insulin expressed when gene replicates with B-galactosidase • Cell undergoes mitosis

  8. STEP 4 • Make functional protein • A and B chains extracted from B-galactosidase and purified • Mixed together • Reconnected in disulfide cross • Then made suitable for injection

  9. Other Human Proteins Made with Genetic Engineering • Erythropoetin- treats anaemia in kidney failure patients • Human Growth Hormone- used to treat pituitary dwarfism • Insulin- Diabetes • Interferon- can treat Hepatitis B and C, some cancers, and MS • Factor VIII- treat hemophilia caused by lack of factor VIII

  10. Factor VIII- Ryan White • Hemophiliac who got AIDS from a blood transfusion • Blood donation has risks - HIV and hepatitis • So, GM bacteria replaced blood donation for hemophiliacs • Factor VIII produced synthetically • Helps clot blood (factor)

  11. Human Growth Hormone • Pituitary Dwarfism- treated by Human Growth Hormone • Before: extracted from pituitary glands of corpses • Some problems • Creutzfeldt-Jacob disease (CJD) • Brain disease • Now: GE bacteria • Injection

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