Efficient PCR‐based gene disruption in Saccharomyces strains using intergenic primers

An Erratum has been published for this article in Yeast 19(9) 2002, 803. 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.