RECOMBINANT DNA TECHNOLOGY PART 2 Quiz Essay 2 Topics Molecular Topics: 35 points Emphasis: data...
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Transcript of RECOMBINANT DNA TECHNOLOGY PART 2 Quiz Essay 2 Topics Molecular Topics: 35 points Emphasis: data...
RECOMBINANT DNA TECHNOLOGY
PART 2
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
Topics• Cloning & Finding cloned genes
– Using bacteria
– Using viruses
• Genomic libraries
• Transgenic plants
• Transgenic animals
• Timely topics
Technique: Cloning of specific genes or DNA fragments
+
= restriction enzyme
= ligase
Cut vector
Cut foreign DNA or gene
Paste together
2 different antibiotic resistant genes
Bacteria are now resistant to 2 antibiotics
Bacterial cloning using plasmids
Cut DNA here & insert foreign DNA
Will your transformed colonies with the insert be white or blue?
Summary of cloning DNA into a blue/white selection
vector
• Clone into a virus
• Infect bacteria
• Identify infected bacteria containing gene
– Assume you know the protein sequence
Cloning and finding desired clone
Bacterial clones and finding desired clone
Probe design
Genomic library contains all of the organism’s genome
Expression Vectors
• Sources of proteins
Expression 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 transgenic plants
• 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.
Transgenic Plants
• In 1996: Soybeans resistant to the weed killer of Round Up.