摘要
用内切酶BglⅡ和EcoRⅠ将PVY-cp基因自pGEM-pvy切下,定向插入到经BamHⅠ和EcoRⅠ双酶切的表达质粒pBV220的启动子Pr、Pl的下游,构建了该基因的原核表达载体pBV-pvy。SDS-PAGE凝胶电泳结果表明:PVY-cp基因在大肠杆菌HB101中经温度(42℃)诱导后,可特异地高效表达大小约30KD蛋白。Western印迹杂交进一步鉴定了该表达蛋白确为PVY外壳蛋白。
The coat protein gene was cut down from pGEM-pvy vector using the restriction endonuclease BglⅡ and EcoRⅠ, and reinserted into the expression vector pBV220 cut by BamHⅠ and EcoRⅠ following the promoter Pr?Pl, and the combination expression vector was obtained, named as pBV-pvy The results of SDS-PAGE electrophoresis and Western Blot analysis showed that the PVY-cp gene could express in Ecoli HB101 through temperature induction and the 30KD expression protein was obtained
出处
《山东农业科学》
2003年第1期3-5,共3页
Shandong Agricultural Sciences
基金
国家农业部"948"项目 (序列号为 :2 0 1 0 47)
山东省科技攻关项目 (编号 :省 99-0 5)。
