甘蓝SRK的S域及SCR酵母表达载体的构建及其相互作用区段的研究  

Studies on the Construction of S Domain of SRK and SCR Gene's Yeast Expression Vectors and Their Interaction in Yeast Two-hybrid System

作　　者：彭一波[1] 朱利泉[1] 杨红[1] 薛丽琰[1] 罗兵[1] 吴志刚[1] 黄丹[1] 杨昆[1] 高启国 李成琼 任雪松 王小佳 

机构地区：[1]西南大学农学与生物科技学院,重庆400716 [2]重庆市蔬菜学重点实验室,重庆400716

出　　处：《园艺学报》2011年第8期1479-1488,共10页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(30971849);高校基本科研业务专项基金项目(XDJKC109)

摘　　要：为了深入研究S位点受体激酶（SRK）和S位点富含半胱氨酸蛋白（SCR）甘蓝自交不亲和（self-incompatibility,SI）信号传导的雌雄决定因子的相互作用机制,以自交不亲和性高的结球甘蓝为材料,采用酵母双杂交体系,构建pGADT7-eSRKs和pGBKT7-SCRs重组载体,并分别转化到Y187和Y2HGold酵母中,通过SD/-Ade-His-Trp-Leu平板上菌落的形成,PCR以及x-α-gal显色反应鉴定转化到酵母中的两个重组质粒相互作用。结果表明,SRK S域的SRK1及SRK4分别与SCR2相互作用,且初步显示其相互作用区域为SRK第1个外显子的第16~421bp的片段和SCR第1876~2068bp的片段。这既为SRK-SCR相互作用提供了具体证据,也为甘蓝自交不亲和性分子机理的深入研究提供了新内容。For further study on the interaction of SRK-SCR and between these two determinant factors,Brassica oleracea with high self-incompatibility was used as research materials to construct a group of pGADT7-eSRKs and pGBKT7-SCRs recombinant vectors that were subsequently transformed accordingly into Y187 and Y2HGold yeast.The interaction between these two recombinant plasmids transformed into the yeasts were tested by SD/-Ade-His-Trp-Leu fungal colony formation of plates,PCR and x-α-gal color reactions.The results showed that there existed two kinds of interactions among all interaction assays,which preliminary indicated that their interaction region span 16-421 bp fragment of the first exon of SRK and 1 876-2 068 bp fragment of SCR.These results reconfirm the SRK-SCR interaction,adding some insights into the mechanism of selt incompatibility in Brassica.

关 键 词：甘蓝 SRK eSRKs基因 SCRs基因 pGADT7载体 pGBKT7载体 酵母双杂交体系 

分 类 号：S635[农业科学—蔬菜学]