Bimolecular fluorescence complementation low analysis system for in vivo detection of protein-protein interaction in Saccharomyces cerevisiae

被引：152
作者：
Sung, Min-Kyung
Huh, Won-Ki [1 ]
机构：
[1] Seoul Natl Univ, Sch Biol Sci, Seoul 151747, South Korea
[2] Seoul Natl Univ, Res Ctr Funct Cellulom, Inst Microbiol, Seoul 151747, South Korea
来源：
YEAST | 2007年 / 24卷 / 09期
关键词：
bimolecular fluorescence complementation; protein-protein interaction; PCR-mediated gene modification; yellow fluorescent protein; subcellullar localization;
D O I：
10.1002/yea.1504
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
The bimolecular fluorescence complementation (BiFC) assay has been widely accepted for studying in vivo detection of protein-protein interactions in several organisms. To facilitate the application of the BiFC assay to yeast research, we have created a series of plasmids that allow single-step, PCR-based G or N-terminal tagging of yeast proteins with yellow fluorescent protein fragments for BiFC assay. By examination of several interacting proteins (Sis1-Sis1, Net1-Sir2, Cet1-Cet1 and Pho2-Pho4), we demonstrate that the BiFC assay can be used to reliably analyse the occurrence and subcellular localization of protein-protein interactions in living yeast cells. The sequences for the described plasmids were submitted to the GenBank under Accession Nos: EF210802, pFA6a-VN-His3MX6; EF210803, pFA6a-VC-His3MX6; EF210804, pFA6a-VN-TRP1; EF210807, pFA6a-VC-TRP1; EF210808, pFA6a-VN-kanMX6; EF210809, pFA6a-VC-kanMX6; EF210810, pFA6a-His3MX6-P-GAL1-VN; EF210805, pFA6a-His3MX6-P-GAL1-VC; EF210806, pFA6a-TRP1-P-GAL1-VN; EF210811, pFA6a-TRP1-P-GAL1-VC; EF210812, pFA6a-kanMX6-P-GAL1-VN; EF210813, pFA6a-kanMX6-P-GAL1-VC; EF521883, pFA6a-Hiis3MX6-P-CET1-VN; EF521884, pFA6a-His3MX6-P-CET1-VC; EF521885, pFA6a-TRP1-P-CET1-VN; EF521886, pFA6a-TRP1-P-CET1-VC; EF521887, pFA6a-kanMX6-P-CET1-VN; EF521888, pFA6a-kanMX6-P-CET1-VC. Copyright (c) 2007 John Wiley & Sons, Ltd.
引用
收藏
页码：767 / 775
页数：9
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