Novel screening system for protein-protein interactions by bimolecular fluorescence complementation in Saccharomyces cerevisiae |
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Authors: | Kojima Takaaki Karasawa Satoshi Miyawaki Atsushi Tsumuraya Takeshi Fujii Ikuo |
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Affiliation: | Department of Biological Science, Osaka Prefecture University, Sakai, Osaka 599-8531, Japan. |
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Abstract: | For high-throughput screening of protein-protein interactions, we have developed a novel yeast screening system using Bimolecular fluorescence complementation (BiFC). Two yeast plasmids, in which genes of heterodimerized peptides LZA and LZB were each fused with those of non-fluorescent half fragments of Kusabira-Green mutant (mKG2), were transformed into a- and α-type yeast, respectively. Mating of them gave a library, which was screened by following green fluorescence resulted from LZA-LZB interaction. The method showed potential ability to detect the positive clones from a model library, in which green-fluorescent and non-fluorescent yeast was mixed in a ratio of 1:675. |
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Keywords: | Saccharomyces cerevisiae Bimolecular fluorescence complementation (BiFC) Protein–protein interactions Flow cytometry Fluorescence screening |
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