recombinant dna technology part 2 topics genomic libraries id a specific clone or sequence within a...
TRANSCRIPT
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RECOMBINANT DNA TECHNOLOGY
PART 2
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Topics• Genomic libraries
• Id a specific clone or sequence within a library
• Transgenic plants
• Transgenic animals
• Timely topics
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Genomic library contains all of the organism’s genome
Probe design
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Bacterial clones and finding desired clone
Probe design
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A Final about vectors : There are many
types
• Small vectors
• Large vectors
• Expression vectors
• Viral vectors
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Transgenic Mice
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transgene
Linear DNA
Non-homologous recombination
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Knockout Mice
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Phenotype of the knockout mouse gives information on protein function
Homozygous knockout
G418gancyclovir
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Applications of DNA technology
• Agriculture
• Drug design
• Genetic testing
• Gene therapy
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Genetic Modified Plants: “GM”
Tobacco with luciferase gene from firefly
Transgenic plant
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Tumor Inducing Plasmid
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Model for making genetically modified (GM) plants
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GM crops
• Bt toxin genes (Bacillus thuringensis)
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Bt Toxin
Cry toxins and cry genes: damages gut of larvae
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GM Plants• Roundup ready plants: herbicide
resistant
• Herbicide known as: Glyphosate
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Glyphosate• Enzyme inhibitor
• Prevents amino acid synthesis
• Prevents weed growth
• Environmental concerns
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Antisense Technology• 5´ C A U G 3´ mRNA
3´ G U A C 5´ Antisense RNA
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Anti-Sense Technology &
FLAVR SAVRTM Tomato
•Anti-sense DNA technology inhibits polygalacturonase (PG)
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The Flavr-Savr Tomato• Ripens on the vine
• Preserves firmness and flavor
• Uses anti-sense technology
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Cloning of a whole organism: Reproductive Cloning
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Somatic Cell Germ Cell
For ex. Mammary cell ovum
SHEEP CELLS
54 27
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Somatic Cell Germ Cell
For ex. Mammary cell ovum
How to process begins:
542754
discarded
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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?”
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Somatic Gene Therapy
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Successes• SCIDS (Severe Combined Immunodeficiency
Disease)
• Head and neck cancers
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But also failures and controversy
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