Solution if any is not audited so use at your

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Unformatted text preview: nt work has demonstrated that mutations in the promoter of p53, a crucial tumor suppressor gene, are found in a large proportion of Li-Fraumeni individuals. 51 : ID 42 er ad lo wn Do Dow nlo ade r ID: 425 1 r ID: 4251 Downloade Test ID: 568 8 : 56 ID st Te (a) Outline a strategy by which you could determine which regions of the p53 promoter are required for gene expression. (2 marks) Deletion (linker scanning mutagenesis)/point mutation (1 mark) and transfection into cell culture for enhancer/reporter assays (1 mark) Dow nlo ade r ID: 425 Down load 1 er ID: 4251 (b) It is also known that the transcription factor NF1 binds as a monomer to a consensus-binding site in the p53 promoter. Briefly describe how you could determine whether a given point mutation in the p53 promoter disrupts direct NF1 binding? (2 marks) 1 425 r ID: nlo ade Dow ID: 568 EMSA using a radiolabeled p53 promoter sequences for the mutant and wild type promoters + protein extracts (1 mark) and an anti-NF1 antibody to show a supershift (1 mark) Alternative answers: ChIP, DnaseI assay (part ma...
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This note was uploaded on 02/08/2013 for the course CSB 349 taught by Professor Moses during the Fall '12 term at University of Toronto.

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