Genetic approaches to the study of protein-protein interactions.

Genetic approaches to the study of protein-protein interactions.
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研究蛋白质-蛋白质相互作用的遗传学方法。

DOI:
10.1006/meth.1999.0861
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发表时间:
1999
期刊:
Methods (San Diego, Calif.)
影响因子:
--
通讯作者:
Appling,DR
Appling,DR
中科院分区:
--
文献类型:
--
作者:
Appling,DR

文献摘要

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This article describes genetic approaches to the study of heterologous protein–protein interactions, focusing on the yeast Saccharomyces cerevisiae as a useful eukaryotic model system. Several methods are described that can be used to search for new interactions, including extragenic suppression, multicopy suppression, synthetic lethality, and transdominant inhibition. Strategies for screening, genetic characterization, and clone identification are described, along with recent examples from the literature. In addition, genetic methods are discussed that can be used to further characterize a newly discovered protein–protein interaction. These include the creation of mutant libraries of a given protein by chemical mutagenesis or polymerase chain reaction, the production of dominant-negative mutants, and strategies for introducing these mutant alleles back into yeast for analysis. Although these genetic methods are quite powerful, they are often just a starting point for further biochemical or cell biological experiments.