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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
Transgenic Mice
transgene
Linear DNA
Non-homologous recombination
Knockout Mice
Phenotype of the knockout mouse gives information on protein function
Homozygous knockout
G418gancyclovir
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 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
Somatic Cell Germ Cell
For ex. Mammary cell ovum
SHEEP CELLS
54 27
Somatic Cell Germ Cell
For ex. Mammary cell ovum
How to process begins:
542754
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