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Yeast. 2002 Mar 15;19(4):319-28.

Efficient PCR-based gene disruption in Saccharomyces strains using intergenic primers.

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  • 1Department of Genetics and Development, Columbia University College of Physicians and Surgeons, New York, NY 10032-2704, USA.

Erratum in

  • Yeast 2002 Jun 30;19(9):803. Kedacche Mehdi [corrected to Keddache Mehdi].

Abstract

Gene disruptions are a vital tool for understanding Saccharomyces cerevisiae gene function. An arrayed library of gene disruption strains has been produced by a consortium of yeast laboratories; however their use is limited to a single genetic background. Since the yeast research community works with several different strain backgrounds, disruption libraries in other common laboratory strains are desirable. We have developed simple PCR-based methods that allow transfer of gene disruptions from the S288C-derived strain library into any Saccharomyces strain. One method transfers the unique sequence tags that flank each of the disrupted genes and replaces the kanamycin resistance marker with a recyclable URA3 gene from Kluyveromyces lactis. All gene-specific PCR amplifications for this method are performed using a pre-existing set of primers that are commercially available. We have also extended this PCR technique to develop a second general gene disruption method suitable for any transformable strain of Saccharomyces.

Copyright 2002 John Wiley & Sons, Ltd.

PMID:
11870855
[PubMed - indexed for MEDLINE]
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