Cellular and moleculr biological study of self-incompatibility in plants

为明确芸苔属植物自交不亲和系统的分子机制，本项目拟对推测与s -locus有关的基因进行结构和功能分析。芸苔属植物自交不亲和相关基因分析。从campestris b . S8和S12菌株的柱头cDNA文库中分离到编码s位点糖蛋白（SLG）和s受体激酶（SRK）的克隆，并对其序列进行了分析。此外，还克隆了SLG12、SLG9和SRK9的基因组DNA，并对其序列进行了分析。SLG反义基因的转化。利用农杆菌介导系统将SLG8的反义结构转化为一株b.s rapa。在获得的转化中，反义基因的作用打破了自交不亲和性。这种基因通过杂交积极地遗传给下一代。SRK激酶结构域的表达。SRK的胞内结构域（激酶结构域）以GST融合蛋白的形式表达。在体外系统中表达的蛋白显示了自磷酸化的活性。

To calrify the molecular mechanism of self-incompatibility system of Brassica, this project aimed the analysis of the structures and functions of genes presumed to be associated with the S-locus.1.Analysis of genes associated with self-incompatibility of Brassica.From the cDNA libraries of stigma of B.campestris S8 and S12 strains, clones encoding SLG (S-locus glycorotein) and SRK (S-receptor kinase) were isolated and their siquences were established. Furthermore, the genomic DNA of SLG12, SLG9 and SRK9 were also cloned ANd their sequences were analyzed.2.Transformation of the antisense gene of SLG.A antisense constract for SLG8 was transformed to a strain of B.rapa with Agrobacterium-mediated system. In a transoformant obtained, the self-incompatibility was broken down with the action of the antisense gene. The gene was actively inherited to hte next generation through the crossing.3.Expression of the kinase domain of SRK.The inner cellular domain of SRK,kinase domain, was expresses as a fusion protein with GST.The expressed protein rvealted the activity of auto-phosphorylation in an in-vitro system.