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S位点受体激酶(SRK)和S位点富含半胱氨酸蛋白/S位点蛋白11(SCR/SP11)分别为甘蓝自交不亲和(self-incom patibility,SI)信号传导的雌雄决定因子。为了深入研究两者的作用机理和进行人工调控,本研究以结球甘蓝ZQ为材料,利用pET.NusA融合蛋白表达系统,将包含SRK胞外域和跨膜域的mSRK蛋白和SCR蛋白在大肠杆菌BL21中融合表达,经SDS-PAGE电泳检测表达出融合蛋白大小分别约为116kD和74kD。进一步将两融合蛋白进行体外相互作用的检测,结果表明mSRK与SCR蛋白能够相互结合形成稳定的复合体,这为下一步实现SRK-SCR复合体聚合与解离的人为调控提供了技术平台。
S-site receptor kinase (SRK) and S-site-rich cysteine protein / S site protein 11 (SCR / SP11) were respectively expressed in self-incompatible (SI) Male and female decision factor. In order to further study the mechanism and regulation of the two, we studied the expression of mSRK protein and SCR protein containing SRK ectodomain and transmembrane domain by using pET.NusA fusion protein expression system in cabbage ZQ. BL21 fusion protein by SDS-PAGE electrophoresis showed that the size of the fusion protein were about 116kD and 74kD. Furthermore, the in vitro interaction between the two fusion proteins was tested. The results showed that the mSRK and SCR proteins could form a stable complex with each other, which provided a technological platform for the next step to realize the human regulation of SRK-SCR complex polymerization and dissociation.