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.