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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.
ID 42 er
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
ade r ID:
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
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.
- Fall '12