BISC 219/2009:Creating a Transgenic Organism

At the end of this semester long series you will have mastered the following skills and concepts using both microorganisms and a multicellular organism.

Experimental Objectives:
In this semester long experiment you will:
 * 1) Master aspectic technique and tissue culture;
 * 2) Use the Agrobacterium system to introduce a foreign gene, the β -glucuronidase gene (gusA) of E. coli, into the cells of tobacco;
 * 3) Take advantage of the totipotency of plant cells and use these transformed cells to regenerate genetically engineered plants;
 * 4) Confirm that the regenerated plants are transformed by
 * 5) measuring the activity of the introduced enzyme, β–glucuronidase (GUS) spectrophotometrically and histochemically;
 * 6) using PCR to test directly for the presence of the introduced genes.

Background Transgenic Plants: Media Recipes Lab 1: Creating the Transgenic Plant: Day 1 Outside of Lab in the first week: Creating the Transgenic Plant: Days 3-5 In Lab 4: Creating the Transgenic Plant: Week 4-5 In Lab 7-8: Creating the Transgenic Plant: Week 8 Lab 8:GUS by histochemistry Lab 8:Structural Evidence for Transgenic Plants: DNA extraction, PCR Lab 9: RE digestion, Agarose gel electrophoresis of the PCR product Lab 10:Leaf Extract Preparation for GUS Spectrophotometric Assay Lab 10:Spectrophotometric Assay for GUS activity Lab 10:Calculations Lab 11: Analyzing the all the data on the putative transgenic and control plants