Module-based construction of plasmids for chromosomal integration of the fission yeast Schizosaccharomyces pombe.

ABSTRACT: Integration of an external gene into a fission yeast chromosome is useful to investigate the effect of the gene product. An easy way to knock-in a gene construct is use of an integration plasmid, which can be targeted and inserted to a chromosome through homologous recombination. Despite the advantage of integration, construction of integration plasmids is energy- and time-consuming, because there is no systematic library of integration plasmids with various promoters, fluorescent protein tags, terminators and selection markers; therefore, researchers are often forced to make appropriate ones through multiple rounds of cloning procedures. Here, we establish materials and methods to easily construct integration plasmids. We introduce a convenient cloning system based on Golden Gate DNA shuf

SUBMITTER: Kakui Y 

PROVIDER: S-EPMC4632507 | biostudies-literature | 2015 Jun

REPOSITORIES: biostudies-literature