CCHC型锌指结构蛋白OsZFP参与调控水稻Oryza sativa侧根的生长发育,但其相关互作蛋白及调控机制未知.以水稻'日本晴''Nipponbare'为试验材料,克隆了OsZFP基因,利用EcoRI和SalI酶切位点构建酵母双杂交钓饵表达载体pGBKT7+OsZFP,验证该表达载体对酵母菌株Y2H无毒性及报告基因自激活现象;采用酵母双杂交技术,从已构建的水稻cDNA文库中筛选到1个阳性互作蛋白,经美国国家生物技术信息中心数据库(NCBI)同源性比对,鉴定为含有T-complex polypeptide 1(TCP-1)结构域的分子伴侣蛋白第7个亚基(Os06g0687700),命名为chaperonin containing TCP-1 eta subunit(CCT-eta),进而通过酵母一对一回复验证该互作蛋白.基于CCT-eta亚基与OsZFP蛋白互作,推测分子伴侣蛋白亚基CCT-eta可能参与调控水稻侧根的生长发育.%OsZFP, a CCHC-type zinc finger protein, has reportedly partic ipated in lateral root development in rice. To determine the molecular mechanism and its interacting proteins in a regulated network, the OsZFP gene from Oryza sativa. 'Nipponbare' was cloned, and then a yeast bait vector, pGBKT7 + OsZFP, was con-structed by EcoRI and SalI for cDNA library screening of rice. Results showed no toxicity or autoactivation with transformation of the Y2H strain containing the pGBKT7+ OsZFP bait vector. In a yeast two-hybrid assay, a positive interacted protein was obtained and confirmed from the cDNA library. Sequencing and homologous alignment from the National Center for Biotechnology Information (NCBI) showed that the interacting protein (named chaperonin containing a T-complex polypeptide 1 (TCP-1) eta subunit, CCT-eta) was the eta subunit of the chaperone which contained a TCP-1 domain. Based on the interaction between OsZFP and the CCT-eta subunit, it was speculated that the CCT-eta subunit of the chaperone protein may be involved in controlling lat-eral root development in rice.
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