ProblemSet4 answers

ProblemSet4 answers - BIMM100 Fall 2009 Problem Set 4 1....

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BIMM100 Fall 2009 Problem Set 4 1. You want to knockout the Finger gene in yeast. a. If the gene is non-essential, how will you select for cells that contain your knockout? You will have a KanMX cassette specific for your gene of interest. You will then select with G418, which will kill all the cells without KanMX b. Unfortunately, it turns out the Finger gene is essential. i. What does this mean and why is it a problem for your experiment? It means the cell is not viable without Finger. This is a problem because yeast are haploid and without that copy the cell will die and thus you will be unable to study phenotypes in cells lacking Finger ii. Describe another way to test for the function of the Finger gene. You can use a regulatable promoter to control Finger expression. For example, the galactose promoter controlling Finger will make the Finger gene in the presence of galactose. When you switch the cells to glucose, txn will be turned off and you can study the cell in the absence of Finger. 2. True or False? A dominant negative mutant removes the wild type copy of that same protein from the cell. Explain. False.
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This note was uploaded on 02/07/2010 for the course BILD 661039 taught by Professor Mel during the Fall '09 term at UCSB.

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ProblemSet4 answers - BIMM100 Fall 2009 Problem Set 4 1....

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