甘蓝SCR识别与结合SRK胞外域核心编码区DNA序列的酵母双杂交检测  

Identifications of DNA Sequences Encoding Key Region of SCR Interacting with SRK Extracellular Domain by Using Yeast Two-Hybrid System

作　　者：薛丽琰[1] 罗兵[1,2] 朱利泉[1] 杨永军[1] 张贺翠[1] 常登龙[1] 陈松[1] 彭一波[1] 杨红[1] 曾静[1] 杨昆[1] 高启国 李成琼 任雪松 王小佳 

机构地区：[1]西南大学植物生理生化实验室,重庆400716 [2]常熟理工学院生物与食品工程学院,江苏常熟215500 [3]重庆市蔬菜学重点实验室,重庆400716

出　　处：《作物学报》2012年第9期1583-1591,共9页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(30971849);重庆市自然科学基金重点项目(cstc2012jjB80010)资助

摘　　要：SCR是芸薹属自交不亲和性的雄性决定因子。为研究SCR与SRK之间相互作用的核心区段，通过构建结球甘蓝的包含系列不同长度的SCR的cDNA序列的pGBKT7融合载体，利用酵母双杂交系统检测SCR与SRK之间的相互作用。结果显示，构建的载体均未出现自激活现象，所获得SCR全长与其相对应SRK胞外域具有相互作用，且相互作用核心编码区位于SCR编码基因的第97～186bp处。实验结果还显示，此单倍型的SCR信号肽剪切位点及相邻的几个氨基酸残基对相互作用实验结果有干扰作用。以上结论为包括甘蓝在内的芸薹属自交不亲和机制的研究提供了新的实验依据。The S-locus cystein-rich protein （SCR） is the male-determining factor of self-incompatibility in Brassica. In this study, the SCR fragments with different lengths amplified from Brassica oleracea L. were ligated with pGBKT7 to construct recombi- nant bait plasmids, which were then transformed into yeast Y2HGold cells for detecting their interaction with the S-locus receptor kinase extracellular domain （eSRK） in Brassica oleracea by using the yeast two-hybrid system. The results showed that recombi- nant vectors were not activated autonomously. The full length SCR could interact with eSRK, and the core region in the SCR was located between 97 and 186 bp. Moreover, the result also indicated that the splicing site of signal peptides of this haplotype SCR and its several adjacent amino acid residues could affect the interaction. These conclusions add some novel insights into the mechanism research of self- incompatibility in Brassica.

关 键 词：甘蓝 自交不亲和 S位点富含半胱氨酸蛋白(SCR) 酵母双杂交 

分 类 号：S635[农业科学—蔬菜学]