TY - CHAP
T1 - Protocols for screening for binding partners of CCN proteins
T2 - Yeast two-hybrid system
AU - Hoshijima, Mitsuhiro
AU - Hattori, Takako
AU - Takigawa, Masaharu
N1 - Publisher Copyright:
© Springer Science+Business Media New York 2017.
PY - 2017
Y1 - 2017
N2 - 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.
AB - 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.
KW - Bait protein
KW - CCN2 interactive proteins
KW - Prey protein
KW - Reporter gene
KW - Yeast two-hybrid system
KW - cDNA library
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U2 - 10.1007/978-1-4939-6430-7_15
DO - 10.1007/978-1-4939-6430-7_15
M3 - Chapter
C2 - 27734374
AN - SCOPUS:84991720112
T3 - Methods in Molecular Biology
SP - 145
EP - 154
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -