紫云英类受体蛋白激酶Nip43靶蛋白的筛选与初步鉴定
Screening and identification of the interacting protein with a RLKs protein Nip43 in Astragalus sinicus

以紫云英类受体蛋白激酶Nip43的S-TKs结构域构建诱饵载体，通过酵母双杂交技术，筛选获得17个候选的阳性克隆。经过测序分析和Blast比对，最终确定了1个互作蛋白EPN。EPN蛋白是网格蛋白聚合反应中的配体蛋白，可调控网格蛋白的聚合反应。通过实验验证显示，靶蛋白EPN与Nip43蛋白互作最强的区域为ENTH/ANTH结构域。Real-time qPCR检测表明，epn基因参与了根瘤菌早期结瘤过程。
The S-TKs domain of Astragalus sinicus gene nip43 was cloned into pGBKT7 and used as a bait plasmid.17 original positive candidate clones were obtained with yeast two-hybrid screening method.One positive clone encoding an EPN protein out of importance was identified through sequencing the cDNA inserts.EPN is an adaptor in the polymerization of clathrins.The critical interaction domain of EPN with Nip43 was determined as ENTH/ANTH domain.Results of real-time qPCR showed that epn gene was involved in the early process of root nodulation.