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Design and Utility of Temperature-Sensitive Gal4 Mutants for Conditional Gene Expression in Drosophila

Mondal, Kajari and VijayRaghavan, K and Varadarajan, Raghavan (2007) Design and Utility of Temperature-Sensitive Gal4 Mutants for Conditional Gene Expression in Drosophila. In: Fly, 1 (5). pp. 282-286.

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Abstract

Temperature-sensitive (ts) mutants are valuable tools to study the function of essential genes in vivo. Despite their widespread use, little is known about mechanisms responsible for the temperature-sensitive (ts) phenotype, or of the transferability of ts mutants of a specific gene between organisms. Since ts mutants are typically generated by random mutagenesis it is difficult to isolate such mutants without efficient screening procedures. We have recently shown that it is possible to obtain ts mutants at high frequency by targeted mutations at either predicted, buried residues important for protein stability or at functional, ligand binding residues. The former class of residues can be identified solely from amino acid sequence and the latter from Ala scanning mutagenesis or from a structure of the protein:ligand complex. Several ts mutants of Gal4 in yeast were generated by mutating both categories of residues. Two of these ts mutants were also shown to result in tight and rapid ts reporter gene-expression in Drosophila when driven by either the elav or GMR promoters. We suggest possible mechanisms that might be responsible for such transferable ts phenotypes and also discuss some of the limitations and difficulties involved in rational design of ts mutants.

Item Type: Journal Article
Additional Information: Copyright of this article belongs to Landes Bioscience.
Department/Centre: Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited: 25 Aug 2008
Last Modified: 28 Feb 2010 11:28
URI: http://eprints.iisc.ernet.in/id/eprint/15619

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