【摘要】 以原核表达载体pGEX-4T-1为基础,通过PCR、酶切与连接构建了含长喙田菁(Sesbania rostrata)植物螯合肽合成酶SrPCS1 cDNA开放阅读框序列的GST-SrPCS1原核表达载体pAM34,并将其转化表达菌株BL21(DE3),在28℃,用0.1 mmol/L IPTG诱导融合蛋白的表达,通过Western blotting鉴定融合蛋白,将SDS-PAGE分离得到的融合蛋白质免疫新西兰白兔,通过Western点杂交对免疫血清的效价进行检测。研究结果表明:通过大肠杆菌表达出GST-SrPCS1融合蛋白,免疫血清的效价为1∶2 000,制备的血清有较高的活性与特异性。更多还原

【Abstract】 The prokaryotic expressing vector pAM34 containing Sesbania rostrata phytochelatin synthase1 open reading frame was constructed on the base of pGEX-4T-1 and transferred into E.coli BL21(DE3),then the fused protein GST-SrPCS1 was induced by 0.1mmol/L at 28℃,identified by western blotting and the rabbit was immunized with the purified protein.The results suggested that the fusion protein GST-SrPCS1 was expressed and the titer of the anti-serum was about 1∶2000,indicating that the polyclonal anti-AtPCSl-His serum has high activity and speciality.更多还原