百脉根结瘤因子受体NFR5及其相互作用蛋白的研究  

Proteins interacted with NFR5 involved in nodulation signaling transduction in Lotus japonicus

作　　者：陈春芬[1] 朱辉[1] 杨绮[1] 朱浩[1] 张忠明[1] 

机构地区：[1]华中农业大学农业微生物学国家重点实验室,湖北武汉430070

出　　处：《中国油料作物学报》2007年第4期372-376,共5页Chinese Journal of Oil Crop Sciences

基　　金：国家自然科学基金(30570056)

摘　　要：以豆科植物百脉根的mRNA为模板,利用RT-PCR方法,扩增到1.0kb结瘤因子受体5(nod factorreceptor NFR5)蛋白激酶结构域的DNA片段(nfr5-pk),并将该片段克隆到酵母双杂交BD载体pGBKT7上。通过酵母双杂交技术,筛选接种根瘤菌的百脉根AD-cDNA酵母双杂交文库,从文库中分离到与nfr5-pk相互作用的221个阳性克隆。从这些阳性克隆酵母中分离AD质粒经反复验证,对26个确定阳性克隆的外源片段进行测序和同源性分析。通过NCBI数据库比对分析鉴定出小G蛋白Rop6等6个与NFR5-PK相互作用的不同蛋白,为进一步研究NFR5基因的功能和调控机制提供了材料。NFR5 is involved in signal perception and transduction of Nod factor produced by rhizobia in Lotus japonicus. In this research, by using yeast two hybrid approach, AD-cDNA library of L. japonicus was constructed and 221 positive clones were idendified by using the protein kinase domain of NFR5 （nfr5- pk） as a bait at pre-screening. After retranformed and X-Gal assay, 26 positive clones were confirmed. Six kinds of proteins including small G-Protein Rop6 were obtained after sequencing and blast search. These proteins proposed to be participated in function regulation NFRS.

关 键 词：酵母双杂交 NFR5 小G蛋白Rop6 百脉根 

分 类 号：Q78[生物学—分子生物学]