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 G418 gancyclovir

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.