BISC 219/2009:Creating a Transgenic Organism
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At the end of this semester long series you will have mastered the following skills and concepts using both microorganisms and a multicellular organism.
Schedule of Experiments
Lab # | Dates | Activity |
---|---|---|
1 | 9/2 - 9/8 | set-up of plant transformation |
Independently | 9/5 - 9/13 | transfer of explants to selective medium; induce shoots |
5 | 9/30 - 10/6 | transfer shoots to root inducing medium |
7 | 10/15 - 10/21 | transfer plantlets to soil |
9 | 11/12 - 11/18 | GUS activity assays; set up PCR reactions |
10 | 11/19 - 11/25 | Gel electrophoresis of PCR product |
11 | 12/9 | Plant Lab Report Due for all students |
Experimental Objectives:
In this semester long experiment you will:
- use the Agrobacterium system to introduce a foreign gene, the β -glucuronidase gene (gusA) of E. coli, into the cells of tobacco
- take advantage of the totipotency of plant cells and use these transformed cells to regenerate genetically engineered plants
- confirm that the regenerated plants are transformed by
- measuring the activity of the introduced enzyme, β–glucuronidase (GUS) spectrophotometrically and histochemically
- PCR to test directly for the presence of the introduced genes and
- become adept at sterile technique.