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Yeast. 2011 Sep;28(9):673-81. doi: 10.1002/yea.1895. Epub 2011 Aug 8.

Application of the FLP/FRT system for conditional gene deletion in yeast Saccharomyces cerevisiae.

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1
Laboratory of Cell Biology, NHLBI, NIH, Bethesda, MD 20892, USA. parky3@mail.nih.gov

Abstract

The yeast Saccharomyces cerevisiae has proved to be an excellent model organism to study the function of proteins. One of the many advantages of yeast is the many genetic tools available to manipulate gene expression, but there are still limitations. To complement the many methods used to control gene expression in yeast, we have established a conditional gene deletion system by using the FLP/FRT system on yeast vectors to conditionally delete specific yeast genes. Expression of Flp recombinase, which is under the control of the GAL1 promoter, was induced by galactose, which in turn excised FRT sites flanked genes. The efficacy of this system was examined using the FRT site-flanked genes HSP104, URA3 and GFP. The pre-excision frequency of this system, which might be caused by the basal activity of the GAL1 promoter or by spontaneous recombination between FRT sites, was detected ca. 2% under the non-selecting condition. After inducing expression of Flp recombinase, the deletion efficiency achieved ca. 96% of cells in a population within 9 h. After conditional deletion of the specific gene, protein degradation and cell division then diluted out protein that was expressed from this gene prior to its excision. Most importantly, the specific protein to be deleted could be expressed under its own promoter, so that endogenous levels of protein expression were maintained prior to excision by the Flp recombinase. Therefore, this system provides a useful tool for the conditional deletion of genes in yeast.

PMID:
21823166
PMCID:
PMC3169912
DOI:
10.1002/yea.1895
[Indexed for MEDLINE]
Free PMC Article
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