A yeast three-hybrid method to clone ternary protein complex components

Jie Zhang, Susan Lautar

Research output: Contribution to journalArticle

Abstract

The yeast two-hybrid system is a powerful technique to detect binary protein interactions. In order to facilitate ternary complex analysis, we have developed the yeast three-hybrid system. Using epidermal growth factor (EGF) receptor, Grb2 and Sos proteins as an example, we demonstrated the use of the three-hybrid system to detect formation of a three-component complex, i.e., the Grb2-mediated interactions between EGF receptor and Sos. We then used the three-hybrid system to screen a library with Gal4-DNA binding domain -EGF receptor and Gal4 activation domain-Sos fusion proteins and isolated a plasmid encoding Grb2. Thus, the three-hybrid system is useful for characterizing and cloning components of ternary protein complexes.

Original languageEnglish (US)
Pages (from-to)68-72
Number of pages5
JournalAnalytical Biochemistry
Volume242
Issue number1
DOIs
StatePublished - Nov 1 1996
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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