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

 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 ~ 421 bp的片段和SCR第1 876 ~ 2 068 bp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。

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

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.