Bimolecular fluorescence complementation low analysis system for in vivo detection of protein-protein interaction in Saccharomyces cerevisiae
Bimolecular fluorescence complementation low analysis system for in vivo detection of protein-protein interaction in Saccharomyces cerevisiae
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DOI：
10.1002/yea.1504
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发表时间：
2007-09-01
期刊：
影响因子：
2.6
通讯作者：
Huh, Won-Ki
中科院分区：
文献类型：
--
作者：
Sung, Min-Kyung;Huh, Won-Ki
双分子荧光互补(BIFC)技术已被广泛应用于多种生物体内蛋白质-蛋白质相互作用的研究。为了促进BIFC分析在酵母研究中的应用，我们创造了一系列的质粒，允许单步、基于PCR的G端或N端标记带有黄色荧光蛋白片段的酵母蛋白用于BIFC分析。通过对几个相互作用的蛋白质(Sis1-Sis1，Net1-Sir2，CET1-CET1和Pho2-Pho4)的检测，我们证明了BIFC法可以可靠地用于分析活的酵母细胞中蛋白质-蛋白质相互作用的发生和亚细胞定位。所描述的质粒序列已提交GenBank，登录号为：EF210802，pFA6a-VN-His3MX6；EF210803，pFA6a-VC-His3MX6；EF210804，pFA6a-VN-TRP1；EF210807，pFA6a-VC-TRP1；EF210808，pFA6a-P-GAL1-VN；EF210809，pFA6a-VC-VC-kanMX6；EF210810，pFA6a-His3MX6-P-GAL1-VN；EF210805，pFA6a-His3MX6-P-GAL1-VN；EF210806，pFA6a-P1-GAL1-VN；EF21011，pFA6a-GAP-1-VN；EF210810，pFA6a-His3MX6-P-GAL1-VN；EF210805，pFA6a-His3MX6-P-GAL1-VN；EF210806，pFA6a-P1-GAL1-VN；EF21011，pFA6a-GALP-GANMX6；EF210805，pFA6a-His3MX6-P-GAL1-VN；EF210805，pFA6a-His3MX6-P-GAL1-VN；EF210806，pFA6a-P1-GAL1-VN；EF21011，pFA6a-GAP-1-VN；EF210811，pFA6a-His3MX6-P-GAL1-VN；EF210805，pFA6a-His3MX6-P-GAL1-VN；EF210805，pFA6a-His3MX6-P-GAL1-VN；EF2EF210813，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。版权所有(C)2007 John Wiley&Sons，Ltd.
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.