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

Yeast. 2007 Sep;24(9):767-75. doi: 10.1002/yea.1504.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Base Sequence
  • DNA, Fungal / chemistry
  • DNA, Fungal / genetics
  • Genetic Vectors / chemistry
  • Genetic Vectors / genetics
  • Luminescent Proteins / chemistry
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism*
  • Microscopy, Fluorescence
  • Molecular Sequence Data
  • Plasmids / chemistry
  • Plasmids / genetics
  • Polymerase Chain Reaction
  • Protein Binding
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Sequence Analysis, DNA


  • Bacterial Proteins
  • DNA, Fungal
  • Luminescent Proteins
  • Saccharomyces cerevisiae Proteins
  • yellow fluorescent protein, Bacteria

Associated data

  • GENBANK/EF210802
  • GENBANK/EF210803
  • GENBANK/EF210804
  • GENBANK/EF210805
  • GENBANK/EF210806
  • GENBANK/EF210807
  • GENBANK/EF210808
  • GENBANK/EF210809
  • GENBANK/EF210810
  • GENBANK/EF210811
  • GENBANK/EF210812
  • GENBANK/EF210813
  • GENBANK/EF521883
  • GENBANK/EF521884
  • GENBANK/EF521885
  • GENBANK/EF521886
  • GENBANK/EF521887
  • GENBANK/EF521888