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Chapter 19 Notes.docx

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Robert Boulianne

Chapter 19 – Applications of Genetic Engineering Insulin from Recombinant DNA Figure Above: To synthesize recombinant insulin, synthetic oligonucleotides encoding the insulin A and B chains were inserted (in separate vectors) adjacent to a cloned E. coli lacZ gene. The recombinant plasmids were transformed into E. coli where the beta gal/insulin fusion was synthesized and accumulated in the cells. Fusion proteins were then extracted from the host cells and the insulin subunits were purified and mixed to produce a functional insulin molecule. Recombinant Approaches for Vaccine Production • Traditional Vaccines – Inactivated vaccines • Prepared from killed samples of bacteria and viruses – Attenuated Vaccines • Weakened or live pathogen which can’t reproduce causing a mild form of the disease • Subunit Vaccines – Cloned surface proteins of pathogenic bacteria and viruses • Protein acts as an antigen and stimulates the immune system to make antibodies against the pathogen • Eg. Guardasil is a subunit vaccine against HPV Herbicide Resistance in Crop Plants • Glyphosate is a herbicide that kills all plants at low concentrations, is biodegradable and is non- toxic to humans – Active herbicide in Roundup – Kills plants by inhibiting chloroplast enzyme EPSP synthase • Necessary for amino acid biosynthesis in both plants and bacteria Movement of Gene into Plants Figure Above: To create glyphosate-resistant transgenic plants, EPSP synthase gene from bacteria is fused to a promoter such as the promoter from the cauliflower mosaic virus. This fusion gene is ligated to a Ti-plasmid vector, the recombinant vector is transformed into an agrobacterium host. Agrobacterium infection of cultured plant cells transfers the EPSP synthase fusion gene into a plant cell chromosome. Cells that acquire the gene are able to synthesize large quantities of EPSP synthase, making them resistant to the herbicide glyphosate. • Leptin – A 167 amino acid peptide hormone found in adipose tissue (fat). – Low levels associated with obesity in mouse models – A “knock-out” mouse was created that could not produce leptin (big mouse in photo) • Injection with leptin can rever
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