期刊
NUCLEIC ACIDS RESEARCH
卷 29, 期 10, 页码 -出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/29.10.e47
关键词
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资金
- Volkswagen Foundation [I/76353]
- Korber Foundation
- European Union [QLG1-1999-00202]
We have developed a novel inducible Cremutant with enhanced recombinase activity to mediate genetic switching events. The protein, designated Cre*PR, is composed of a new Cre mutant at the N-terminus followed by the ligand-binding domain (LBD) of the progesterone receptor (PR). The response to low doses of inducer is significantly enhanced by elongating the C-terminus of the PR LBD from amino acid 891 to 914. The mutant Cre lacks the first 18 amino acids and contains a Val -> Ala substitution at position 336, thereby destroying a cryptic splice donor at the 3'-end of Cre. The latter mutation reduces unwanted background recombinase activity in the absence of the synthetic ligand RU486 by a factor of at least 10 to an almost undetectable level. Thus, the recombinase activity turns out to be inducible by a factor of > 200. We expect Cre*PR to serve as a valuable tool for conditional expression of genes both in vitro and in vivo.
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