RECOMBINANT DNA TECHNOLOGY

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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 organisms
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
Non-homologous recombination
Linear DNA
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Knockout Mice
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G418 gancyclovir
Homozygous knockout
Phenotype of the knockout mouse gives information
on protein function
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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     mRNA3   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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SHEEP CELLS
54
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Somatic Cell Germ Cell For ex. Mammary
cell ovum
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How to process begins
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54
discarded
Somatic Cell Germ Cell For ex. Mammary
cell ovum
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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 lambs 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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