Week5Q_07 - metabolism Why would you use the following...

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Week 5 Questions: 1. Modified from the “Working with Molecular Cell Biology” study guide: Why are the calcium-chelating agents EGTA or EDTA and the enzyme trypsin used to prepare single cell suspensions from cultured epithelial cell monolayers? (What ECM molecules do they affect?) 2. Matching: ____ Have a high recombination efficiency A. Motif ____ Restores the function of another mutated protein B. Intron ____ Related genes diverged from speciation C. Yeast ____ Related genes derived from gene duplication D. Mouse cells ____ Zinc finger E. Second site suppresor ____ Parasitic DNA elements F. Paralog ____ Non-coding region of a gene G. Ortholog ____ Uses fluorescently-labelled DNA sequences to H. Exon visualize them on chromosomes I. SINEs ____ Dependent on polymorphisms created by unequal J. Recovery gene recombination events K. Satellite DNAs L. FISH M. DNA fingerprinting 3. You knockout a specific gene, GLUT2, in the liver to see its effects on glucose
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Unformatted text preview: metabolism. Why would you use the following steps in your procedure? a. Selection of ES cells with G418 b. Selection of ES cells with ganciclovir c. Southern blot using a blood sample from potential heterozygous mice (brown coat progeny) d. Mouse with Cre gene and heterogygous for GLUT2 crossed with a mouse containing the GLUT2 gene flanked with LoxP sites 4. What is the hallmark of retroviral DNA (key unique feature/sequence) and what is its purpose? 5. In outline, how does RNAi work? What are three limitations of this technique? 6. What would you expect to see when overexpressing a dominant negative mutant protein (as compared to wild type or loss of function mutations)? What is an advantage of using dominant negative mutants? Why are most dominant negative mutants of multi-subunit proteins (when would a single subunit dominant negative mutant be possible)?...
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This note was uploaded on 03/26/2008 for the course BIOBM 4320 taught by Professor Vogt,vm during the Spring '07 term at Cornell.

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