bis_104_mid_i_exam_key_b08 - Page 5

bis_104_mid_i_exam_key_b08 - Page 5 - by light microscopy c...

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Page 4 Name_____________________________ 11. It has been hypothesized that the abnormally high rate of cell division seen in cancer cells may be due to the over-expression of specific growth factor receptors on their plasma membranes (e.g., over-expression of the receptor erbB2 is seen in some breast cancers). Using a DNA construct with beta-galactosidase as your reporter gene, design one experiment to test the hypothesis that over-expression of erbB2 is sufficient to convert normal cells and cause them to proliferate at a rate similar to that of cancer cells. Describe the results you would expect to observe if the hypothesis is correct. EXPERIMENT: (6) a. transfect a pure population of normal cells with a DNA construct composed of: promoter – erbB2 gene – Beta gal reporter b. following transfection, monitor cells for (1) proliferation rate, and (2) expression of B-gal
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Unformatted text preview: by light microscopy. c. compare proliferation rate of transfected cells (those expressing B-gal) to that of normal cells (or tumor cells). RESULTS: (4) Cells expressing B-gal (erbB2) divide more frequently than normal cells (the same cell population prior to transfection). Describe one essential positive control for your experiment and provide the expected results . (10) CONTROL: (6) Verify that B-gal positive transfected cells actually co-express more erbB2 by measuring erbB2 protein on the PM directly: treat cells with a labeled antibody specific for erbB2 and quantify label or extract all PM proteins and quantify (e.g., western blot). RESULTS OF CONTROL: (4) Proliferating cells that are B-gal positive do in fact have elevated levels of erbB2 protein in their PMs. Demo...
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