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

Nature Protocols, ISSN: 1750-2799, Vol: 2, Issue: 11, Page: 2996-3000
2007
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Article Description

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. © 2007 Nature Publishing Group.

Bibliographic Details

Gregan, Juraj; Zhang, Chao; Rumpf, Cornelia; Cipak, Lubos; Li, Zhang; Uluocak, Pelin; Nasmyth, Kim; Shokat, Kevan M

Springer Science and Business Media LLC

Biochemistry, Genetics and Molecular Biology

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