甘蓝自交不亲和决定因子的体外表达和相互作用的检测

 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-incompatibility, 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116 kD和74 kD。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。

In vitro Study on the Interactions Between Determinant Factors of Self-incompatibility in Brassica oleracea L. var. capitata L.

Self-incompatibility of Brassica oleracea L. var. capitata L. is determined by the female determinant S-locus receptor kinase(SRK) and the male determinant S-locus cysteine rich protein/S-locus protein 11(SCR/SP11). For further study on mechanism and reagent control of the interaction between the determinant factors, extracellular and transmember domains of SRK(named as mSRK) and open reading frame of SCR were amplified from Brassica oleracea L. cv. ZQ. Then fusion protein of mSRK about 116 kD and SCR-orf about 74 kD were expressed in E. coli BL21 by the pET·NusA fusion expression system and separated by SDS-PAGE electrophoresis. After SCR-orf was incubated with mSRK for 2 hours, the interaction between the two proteins was analyzed; the result shows that mSRK and SCR could combine with each other and form a stable complex.