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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.
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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 design
A Final about vectors : There are many types • Small vectors • Large vectors • Expression vectors • Viral vectors
transgene Non-homologous recombination LinearDNA http://spine.rutgers.edu/cellbio/assets/flash/transgen.htm
G418 gancyclovir Homozygous knockout http://www.bio.davidson.edu/courses/genomics/method/homolrecomb.htm 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
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´ mRNA3´ G U A C 5´ 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
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?”
Successes • SCIDS (Severe Combined Immunodeficiency Disease) • Head and neck cancers