BISC110/F09:Series3
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At the end of this three week series on photosynthesis (Labs 8-10) you will have mastered the following skills and concepts.
Series Three- Photosynthesis-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.
Concepts | Genes/Organisms | Techniques/Skills |
---|---|---|
Effect of random gene insertion in a large genome | Bacterial pathogen: Agrobacterium tumefaciens | Aseptic tissue culture |
Plant totipotency | Tobacco plant: Nicotiana tobacum | DNA Extraction |
Regulation of gene expression | gusA gene from E. coli | PCR |
Genetic Engineering and creation of genetically modified organism | Agarose gel electrophoresis | |
Phenotypic Selection | Transfection & selection by antibiotic resistance;
Enzyme function assays: colorimetric & histochemical |
Experiment 8: Photosynthetic Pigments
Experiment 9: The Hill Reaction
Experiment 10: Variables Affecting the Hill Reaction
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