Protocols for Screening for Binding Partners of CCN Proteins: Yeast Two-Hybrid System.
Protocols for Screening for Binding Partners of CCN Proteins: Yeast Two-Hybrid System.
复制标题
CCN 蛋白结合伴侣的筛选方案：酵母双杂交系统。
DOI：
10.1007/978-1-4939-6430-7_15
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Takigawa M
中科院分区：
文献类型：
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作者：
Hoshijima M;Hattori T;Takigawa M
酵母双杂交筛选是鉴定与感兴趣的蛋白质相互作用的蛋白质的有力方法。CCN 2由四个结构域组成，并且鉴定与CCN 2的各个结构域结合的新蛋白质可以揭示CCN 2的各种功能。为了确定CCN 2相互作用的蛋白质，调节CCN 2的活性，我们进行了基于GAL 4的酵母双杂交筛选与来自软骨细胞系，HCS-2/8的cDNA文库，CCN 2 cDNA用作诱饵。在这一章中，我们详细描述了我们的方法筛选CCN 2的结合伙伴，通过这个系统。该方案也可以应用于其他CCN蛋白。
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.