2002/02/21 by Robert J.D. Reid, Robert J. D. Reid, Ivana Sunjevaric +3
Biochemistry, Genetics and Molecular Biology · Agricultural and Biological Sciences · #Fungal and yeast genetics research #Microbial Metabolic Engineering and Bioproduction #Fermentation and Sensory Analysis
paper · doi:10.1002/yea.817
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.