利用分裂泛素酵母双杂交技术钓取小麦TaSC互作蛋白质

Fishing TaSC Interacting Protein in Wheat Using Split Ubiquitin Yeast Two Hybrid System

TaSC(Triticum asetivum L.salt-tolerance related gene,Gen Bank登录号为AY956330)是从小麦耐盐突变体RH8706-49中克隆的高盐诱导表达的耐盐相关基因。以该基因编码的蛋白质TaSC为诱饵,运用分裂泛素酵母双杂交技术从小麦cDNA表达文库中钓取互作蛋白质,筛选到一个编码小麦未知功能蛋白质的基因(GenBank登录号为AK336035),命名为TaSCIP1(TaSC interaction protein 1)。双分子荧光互补(BiFC)实验证实TaSCIP1与TaSC存在互作。该互作蛋白的分离有利于进一步研究TaSC基因的耐盐机制。

The TaSC （Triticum asetivum L. salt-tolerance related gene, GenBank accession number AY956330） has been cloned from wheat salt tolerant mutant RH 8706-49, which is induced by high salinity. To fish interacting proteins of this gene, we used TaSC protein as bait to hybrid with the wheat cDNA expression library using the split ubiquitin yeast two hybrid system in this experiment. A novel wheat gene （GenBank accession number AK336035） encoding a protein with unknown function was isolated, which was designated TaSCIP1 （TaSC interaction protein 1）. Bimolecular fluorescence complementation （BiFC） experiment con- firmed the interaction between proteins TaSCIP1 and TaSC. The separation of TaSC-interacting protein is beneficial to disclose the mechanism of salt tolerance gene TaSC.