Designer deletion strains derived fromSaccharomyces cerevisiae S288C: A useful set of strains and plasmids for PCR-mediated gene disruption and other applications

Carrie Baker Brachmann(Johns Hopkins University), Adrian Davies(Johns Hopkins University), Gregory J. Cost(Johns Hopkins University), Emerita M. Caputo(Johns Hopkins University), Joaquim Li(University of California, San Francisco), Philip Hieter(Johns Hopkins University), Jef D. Boeke
Cited by 3,546

Abstract

A set of yeast strains based on Saccharomyces cerevisiae S288C in which commonly used selectable marker genes are deleted by design based on the yeast genome sequence has been constructed and analysed. These strains minimize or eliminate the homology to the corresponding marker genes in commonly used vectors without significantly affecting adjacent gene expression. Because the homology between commonly used auxotrophic marker gene segments and genomic sequences has been largely or completely abolished, these strains will also reduce plasmid integration events which can interfere with a wide variety of molecular genetic applications. We also report the construction of new members of the pRS400 series of vectors, containing the kanMX, ADE2 and MET15 genes.


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