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Wellesley College-BISC110 Introduction to Cell Biology-Fall 2009

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:

  1. use the Agrobacterium system to introduce a foreign gene, the β -glucuronidase gene (gusA) of E. coli, into the cells of tobacco
  2. take advantage of the totipotency of plant cells and use these transformed cells to regenerate genetically engineered plants
  3. confirm that the regenerated plants are transformed by
    1. measuring the activity of the introduced enzyme, β–glucuronidase (GUS) spectrophotometrically and histochemically
    2. PCR to test directly for the presence of the introduced genes and
  4. 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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