Screening and identification of target proteins interacting with NORK (nodulation receptor kinase) of Astragalus sinicus | 紫云英结瘤受体激酶靶蛋白的筛选与鉴定
2013
Wang Ning, Huazhong Agricultural University, Wuhan (China), College of Life Science and Technology | Li Yixing, Huazhong Agricultural University, Wuhan (China), College of Life Science and Technology | Liu Yan, Huazhong Agricultural University, Wuhan (China), College of Life Science and Technology
中国人. 利用酵母双杂交技术，以结瘤受体激酶（NORK）蛋白的跨膜结构域（TM）为诱饵，进行了与其相互作用靶蛋白的筛选鉴定。结果获得41个候选阳性克隆，通过测序和在NCBI中进行Blast比对分析，发现其中一个具有重要生物学功能的互作蛋白翻译延伸因子AsEF1-α，并将该蛋白相应的编码基因命名为AsEF1-α。进一步的验证表明靶蛋白AsEF1-α与诱饵互作的关键结构域是EF1-α-Ⅲ。
显示更多 [+] 显示较少 [-]英语. The nodulation receptor kinase (NORK) of Astragalus sinicus mainly contains three functional domains including repetitive sequences of the N-terminal leucine-rich domain (LRR), the central transmembrane domain (TM), and the C-terminal kinase domain (PK), respectively. Using yeast two-hybrid technique and TM domain as the bait protein, its interacting target proteins were screened and identified. 41 original positive clones were obtained. After the inserted cDNA sequencing and Blast analyses in the NCBI database, an interacting target protein with the same biological function as a translation elongation factor protein EF1α was identified. The corresponding protein encoding gene was named AsEF1-α. The further confirmed experiment showed that the key interacted domain of AsEF1-α protein was located in the EF1-α-Ⅲ. These results provide a fundamentally theoretical basis for future in-depth study of the genes’ function and mechanism, and also provide novel information and material for establishing early root nodulation signaling network in legume plant.
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