An improved bimolecular fluorescence complementation tool based on superfolder green fluorescent protein.
An improved bimolecular fluorescence complementation tool based on superfolder green fluorescent protein.
复制标题
一种改进的基于超级文件夹绿色荧光蛋白的双分子荧光互补工具。
DOI：
10.1093/abbs/gmq128
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发表时间：
2011
影响因子：
3.7
通讯作者：
Bin Xia
中科院分区：
文献类型：
--
作者：
Jun Zhou;Jian Lin;C. Zhou;Xiaoyan Deng;Bin Xia
Bimolecular fluorescence complementation (BiFC) has been widely used in the analysis of protein-protein interactions (PPIs) in recent years. There are many notable advantages of BiFC such as convenience and direct visualization of PPI in cells. However, BiFC has one common limitation: the separated non-fluorescent fragments can be spontaneously self-assembled into an intact protein, which leads to false-positive results. In this study, a pair of complementary fragments (sfGFPN and sfGFPC) was constructed by splitting superfolder GFP (sfGFP) between the 214 and 215 amino acid residue, and sfGFPC was mutated by site-directed gene mutagenesis to decrease the signal of negative control. Our results showed that mutations in sfGFPC (sfGFPC(m12)) can effectively decrease the signal of negative control. Thus, we provide an improved BiFC tool for the analysis of PPI. Further, since the self-assembly problem is a common shortcoming for application of BiFC, our research provides a feasible strategy for other BiFC candidate proteins with the same problem.
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影响因子：
15
作者：
Fernandez-Suarez, Marta;Chen, T. Scott;Ting, Alice Y.
通讯作者：
Ting, Alice Y.
影响因子：
2.7
作者：
Shyu, YJ;Liu, H;Hu, CD
通讯作者：
Hu, CD
影响因子：
46.2
作者：
Kerppola TK
通讯作者：
Kerppola TK
影响因子：
4.8
作者：
Hiatt, Susan M.;Shyu, Y. John;Hu, Chang-Deng
通讯作者：
Hu, Chang-Deng