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Nat Protoc. 2007;2(11):2996-3000.

Construction of conditional analog-sensitive kinase alleles in the fission yeast Schizosaccharomyces pombe.

Author information

1
Department of Chromosome Biology, Max F. Perutz Laboratories, University of Vienna, Dr. Bohr-Gasse 1, 1030 Vienna, Austria. juraj.gregan@univie.ac.at

Abstract

Reversible protein phosphorylation is a major regulatory mechanism in a cell. A chemical-genetic strategy to conditionally inactivate protein kinases has been developed recently. Mutating a single residue in the ATP-binding pocket confers sensitivity to small-molecule inhibitors. The inhibitor can only bind to the mutant kinase and not to any other wild-type kinase, allowing specific inactivation of the modified kinase. Here, we describe a protocol to construct conditional analog-sensitive kinase alleles in the fission yeast Schizosaccharomyces pombe. This protocol can be completed in about 3 weeks and should be applicable to other organisms as well.

PMID:
18007635
PMCID:
PMC2957860
DOI:
10.1038/nprot.2007.447
[Indexed for MEDLINE]
Free PMC Article

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