A system for multilocus chromosomal integration and transformation-free selection marker rescue.

ABSTRACT: Yeast integrating plasmids (YIPs) are a versatile tool for stable integration in Saccharomyces cerevisiae. However, current YIP systems necessitate time- and labor-intensive methods for cloning and selection marker rescue. Here, we describe the design, construction, and validation of a new YIP system capable of accelerating the stable integration of multiple expression constructs into different loci in the yeast S. cerevisiae. These 'directed pop-out' plasmids enable a simple, two-step integration protocol that results in a scarless integration alongside a complete rescue of the selection marker. These plasmids combine three key features: a dedicated 'YIPout' fragment directs a recombination event that rescues the selection marker while avoiding undesired excision of the target DNA sequenc

SUBMITTER: Siddiqui MS 

PROVIDER: S-EPMC4270834 | biostudies-literature | 2014 Dec

REPOSITORIES: biostudies-literature