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Bimolecular fluorescence complementation analysis system for in vivo detection of protein–protein interaction in Saccharomyces cerevisiae
Author(s) -
Sung MinKyung,
Huh WonKi
Publication year - 2007
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.1504
Subject(s) - bimolecular fluorescence complementation , protein fragment complementation assay , biology , saccharomyces cerevisiae , yeast , complementation , plasmid , microbiology and biotechnology , protein–protein interaction , biochemistry , gene , phenotype
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 C‐ 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‐His3MX6‐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 © 2007 John Wiley & Sons, Ltd.