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

Transgenic Mice

transgene Linear DNA Non-homologous recombination

Knockout Mice

Phenotype of the knockout mouse gives information on protein function Homozygous knockout G418 gancyclovir

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´ mRNA 3´ G U A C 5´ Antisense RNA

Anti-Sense Technology & FLAVR SAVR TM 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

Somatic CellGerm Cell For ex. Mammary cellovum SHEEP CELLS 54 27

Somatic CellGerm Cell For ex. Mammary cellovum How to process begins: discarded

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