Protocols for screening for binding partners of CCN proteins: Yeast two-hybrid system

Research output: Chapter in Book/Report/Conference proceedingChapter

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Abstract

Yeast two-hybrid screening is a powerful method to identify proteins that interact with a protein of interest. CCN2 consists of four domains, and identification of new proteins that bind to individual domains of CCN2 may reveal a variety of CCN2 functions. To identify CCN2-interactive proteins that regulate CCN2 activity, we carried out GAL4-based yeast two-hybrid screening with a cDNA library derived from a chondrocytic cell line, HCS-2/8, with CCN2 cDNA used as a bait. In this chapter, we describe our methods for screening for CCN2 binding partners by this system in detail. This protocol may be applied to other CCN proteins as well.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages145-154
Number of pages10
DOIs
Publication statusPublished - Jan 1 2017

Publication series

NameMethods in Molecular Biology
Volume1489
ISSN (Print)1064-3745

Keywords

  • Bait protein
  • CCN2 interactive proteins
  • Prey protein
  • Reporter gene
  • Yeast two-hybrid system
  • cDNA library

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Hoshijima, M., Hattori, T., & Takigawa, M. (2017). Protocols for screening for binding partners of CCN proteins: Yeast two-hybrid system. In Methods in Molecular Biology (pp. 145-154). (Methods in Molecular Biology; Vol. 1489). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6430-7_15