Problemset1SP13_000

Gene x is normally expressed in the liver and the

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liver-specific promoter. Gene X is normally expressed in the liver and the pancreas and produces a 15 kb mRNA. To verify that the conditional knock-out worked, you perform a Northern blot with mRNA extracted from the liver, the pancreas and the brain, which you probe with a radioactive probe against the coding sequences of gene X (see next page). Which lane corresponds to which tissue?
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12. You are designing an experiment that requires a knock-out of exon 6 of the FGFR3 gene. On the scheme below, indicate the locations of all necessary elements that will allow the construct to work (Neo, tk, regions of homology, loxP, select the relevant elements). Exon 5 Exon 6 Exon 7 Briefly outline the next steps that need to be taken to obtain a mouse that is heterozygous for the knockout. For example: Step 1. Inject or electroporate the vector into mouse embryonic stem cells Step 2. Select for …. 13. You are studying a different form of autosomal dominant dwarfism, thanatophoric dysplasia (TD). Upon sequencing the cDNA of FGFR3 of the patients with TD, you discover that many of these patients carry a mutation in FGFR3 gene, G370C (conversion of Glycine 370 to Cysteine). You hypothesize that TD is caused by the mutation in FGFR3. Design an experiment in mice that would test whether TD is indeed caused by this mutation. Indicate what animals will you generate, what tissues or cells you will be looking at and how.
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Gene X is normally expressed in the liver and the pancreas...

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