长喙田菁SrPCS1的原核表达及多抗血清的制备  

Prokaryotic Expression of the Recombinant SrPCS1 and Preparation of Polyclonal Anti-serum Against SrPCS1

作　　者：李安明[1] 邓青云[1] 李德华[1] 

机构地区：[1]孝感学院生命科学技术学院,湖北孝感432000

出　　处：《江西农业大学学报》2011年第4期812-816,共5页Acta Agriculturae Universitatis Jiangxiensis

基　　金：湖北省自然科学基金项目(2005ABA083);湖北省重点(培育)学科:农业资源利用(0903)

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。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.

关 键 词：植物螯肽合成酶 原核表达 融合蛋白 抗血清 

分 类 号：Q943.2[生物学—植物学]