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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。
In order to further study the interaction mechanism between male and female determinants of S-site receptor kinase (SRK) and S-site rich self-incompatibility (SI) signaling in cysteine-rich protein The recombinant vectors pGADT7-eSRKs and pGBKT7-SCRs were constructed and transformed into Y187 and Y2HGold yeast respectively by using the yeast two-hybrid system with self-incompatibility of cabbage as raw material. SD / -Ade-His- Colony formation on Trp-Leu plates, PCR, and x-α-gal chromogenic reaction identified the interaction of two recombinant plasmids transformed into yeast. The results showed that SRK1 and SRK4 of SRK S interacted with SCR2 respectively, and the interaction region of SRK1 and SRK4 was initially shown as the 16th ~ 421 bp fragment of the first exon of SRK and the 1876 ~ 2068 bp fragment of SCR. This not only provided concrete evidence for SRK-SCR interaction, but also provided new contents for further research on the molecular mechanism of self-incompatibility of cabbage.