recombinant dna technology part 2 quiz essay 2 topics molecular topics: 35 points emphasis: data...
TRANSCRIPT
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RECOMBINANT DNA TECHNOLOGY
PART 2
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Quiz Essay 2 Topics
Molecular Topics: 35 points
Emphasis: data interpretation, critical thinking with a short answer format
■ Bacterial transformation
■ Plasmid isolation
■ Translation: genetic code table
■ Lac operons: mutational analysis
■ Recombinant DNA technology
Genetics’ Topic 15 points
■ Genetic Fine Structure
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Topics• Cloning & Finding cloned genes
– Using bacteria
– Using viruses
• Genomic libraries
• Transgenic plants
• Transgenic animals
• Timely topics
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Technique: Cloning of specific genes or DNA fragments
+
= restriction enzyme
= ligase
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Cut vector
Cut foreign DNA or gene
Paste together
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2 different antibiotic resistant genes
Bacteria are now resistant to 2 antibiotics
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Bacterial cloning using plasmids
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Cut DNA here & insert foreign DNA
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Will your transformed colonies with the insert be white or blue?
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Summary of cloning DNA into a blue/white selection
vector
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• Clone into a virus
• Infect bacteria
• Identify infected bacteria containing gene
– Assume you know the protein sequence
Cloning and finding desired clone
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Bacterial clones and finding desired clone
Probe design
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Genomic library contains all of the organism’s genome
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Expression Vectors
• Sources of proteins
Expression 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 transgenic plants
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• Bt toxin genes (from Bacillus thuringensis) introduced into corn, making it resistant to the European corn borer.
• Millions acres of corn, cotton, and potatoes bearing Bt toxin are under cultivation.
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Transgenic Plants
• In 1996: Soybeans resistant to the weed killer of Round Up.
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