GENE DISRUPTION WITH PCR PRODUCTS IN SACCHAROMYCES-CEREVISIAE

We describe here the generation of gene disruption constructs using PC R amplification of selectable markers with primers that provide homolo gy to the target gene of interest. We find that regions of homology as short as 38 to 50 bp suffice to mediate homologous recombination in y east. We describe applications of this technology to three specific ye ast genes that would have been difficult to disrupt with current metho ds. By dispensing with the need to either clone the gene of interest o r engineer a standard disruption construct, this method should facilit ate analysis of sequenced genes of unknown function, which will soon i nclude the entire yeast genome.