Molecular Cloning Methods - Santa Monica College

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 organisms genome Probe design Bacterial clones and finding desired

clone Probe des A Final types

about vectors : There are many 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

3 C A U G 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 lambs clothing? Somatic Gene Therapy

Successes SCIDS (Severe Combined Immunodeficiency Disease) Head and neck cancers

But also failures and controversy

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