Rapid, efficient and precise allele replacement in the fission yeast Schizosaccharomyces pombe.

ABSTRACT: Gene targeting provides a powerful tool to modify endogenous loci to contain specific mutations, insertions and deletions. Precise allele replacement, with no other chromosomal changes (e.g., insertion of selectable markers or heterologous promoters), maintains physiologically relevant context. Established methods for precise allele replacement in fission yeast employ two successive rounds of transformation and homologous recombination and require genotyping at each step. The relative efficiency of homologous recombination is low and a high rate of false positives during the second round of gene targeting further complicates matters. We report that pop-in, pop-out allele replacement circumvents these problems. We present data for 39 different allele replacements, involving simple and compl

SUBMITTER: Gao J 

PROVIDER: S-EPMC3954454 | biostudies-literature | 2014 May

REPOSITORIES: biostudies-literature