Journal
FEMS YEAST RESEARCH
Volume 14, Issue 7, Pages 1048-1054Publisher
WILEY-BLACKWELL
DOI: 10.1111/1567-1364.12197
Keywords
selectable marker recycling; gene disruption; plasmid integration; Cre recombinase; yeast
Funding
- Russian Foundation for Basic Research [12-04-32080]
- Program for Molecular and Cell Biology from the Russian Academy of Science
- Russian Scientific Foundation [14-14-00361]
- Russian Science Foundation [14-14-00361] Funding Source: Russian Science Foundation
Ask authors/readers for more resources
Site-specific recombinases are widely used for selectable marker recycling in molecular-genetic manipulations with eukaryotic cells. This usually involves the use of two genetic constructs, one of which possesses a selectable marker flanked by the recombinase recognition sequences, while the other one bears the recombinase gene. Combining the recombinase gene with its recognition sequences in one plasmid is usually avoided, as it may lead to undesirable recombination due to promoter leakage, while the plasmid is maintained in Escherichia coli cells. Here, we describe yeast vectors possessing Cre recombinase genes under control of regulatable yeast promoters and loxP sequences for the in vivo vector backbone excision. The plasmid stability in E.coli is ensured by the presence of an intron in the recombinase gene. Applicability of these vectors was validated by disruptions of the Hansenula polymorpha PMC1 and Saccharomyces cerevisiae HSP104 and PRB1 genes.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available