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RECOMBINANT DNA TECHNOLOGY PART 2 Topics • Genomic libraries • Id a specific clone or sequence within a library • Transgenic plants • Transgenic animals • Timely topics Genomic library contains all of the organism’s genome Probe design Bacterial clones and finding desired clone Probe des A Final about vectors : There are many types • Small vectors • Large vectors • Expression vectors • Viral vectors Transgenic Mice transgen Non-homologous recombination Linear DNA Knockout Mice G418 gancyclovir Homozygous knockout Phenotype of the knockout mouse gives information on protein function Applications of DNA technology • Agriculture • Drug design • Genetic testing • Gene therapy Genetic Modified Plants: “GM” Tobacco with luciferase gene from firefly Transgenic plant Tumor Inducing Plasmid Model for making genetically modified (GM) plants GM crops • Bt toxin genes (Bacillus thuringensis) Bt Toxin Cry toxins and cry genes: damages gut of larvae GM Plants • Roundup ready plants: herbicide resistant • Herbicide known as: Glyphosate Glyphosate • Enzyme inhibitor • Prevents amino acid synthesis • Prevents weed growth • Environmental concerns Antisense Technology • 5´ C A U G 3´ 3´ G U A C 5´ mRNA Antisense RNA Anti-Sense Technology & FLAVR SAVRTM Tomato •Anti-sense DNA technology inhibits polygalacturonase (PG) The Flavr-Savr Tomato • Ripens on the vine • Preserves firmness and flavor • Uses anti-sense technology Cloning of a whole organism: Reproductive Cloning SHEEP CELLS 54 27 Somatic Cell Germ Cell For ex. Mammary cell ovum How to process begins: 54 27 54 discarded Somatic Cell Germ Cell For ex. Mammary cell ovum Now the germ cell contains 54 chromosomes! • The germ is stimulated “electrically”. • Cell division in some instances begins. • Individual is identical to host “DNA” donor. • Aging affects? “Was Dolly a sheep in lamb’s clothing?” Somatic Gene Therapy Successes • SCIDS (Severe Combined Immunodeficiency Disease) • Head and neck cancers But also failures and controversy
