小菜蛾颗粒体病毒PP31蛋白的原核表达及多克隆抗体制备  

Expression and Antibody Preparation of Plutella xylostella Granulovirus PP31

作　　者：王思民[1] 张鸿杰[1] 黎路林[1] 

机构地区：[1]华中师范大学生命科学学院遗传调控与整合生物学湖北省重点实验室,武汉430079

出　　处：《生物技术通报》2013年第3期129-133,共5页Biotechnology Bulletin

基　　金：国家自然科学基金项目(30770088)

摘　　要：杆状病毒PP31是一种磷酸化的DNA结合蛋白。pp31基因存在于所有已完成测序的鳞翅目昆虫杆状病毒。用PCR方法从小菜蛾颗粒体病毒基因组扩增pp31基因,将其克隆至原核表达载体pET28a,转化大肠杆菌BL21(DE3)菌株。经IPTG诱导,在被转化菌株中表达产生出携带6×His标签的PP31蛋白。用ProBondTM树脂纯化的His-PP31融合蛋白大小约29 kD左右。用该融合蛋白免疫新西兰大白兔,获得PlxyGV PP31蛋白抗血清。Western blot分析显示,所获得的抗血清具有较强的特异性和高效价,可应用于PlxyGV PP31蛋白分析。Baculovirus PP31 is a phosphoprotein that could combine with DNA. The pp31 gene presents in all of the genomes of lepidopteran bactdoviruses sequenced to date. In this study, Plutella xylostella pp31 gene was PCR-amplified and cloned into an expression vector pET28a, and transformed into the E. coli BL21 （ DE3 ） . In the transformed bacterial cells, induced by IPTG, a His-tagged PP31 protein with a size of 29 kD was expressed, which was purified using ProbondTM resin. Polyclonal antibodies against PlxyGV PP31 were prepared by immunizing a New Zealand rabbit with the purified hls-tagged PP31. Western blot analysis showed that the antiserum had high titer and specificity and could be used in study of PP31.

关 键 词：小菜蛾颗粒体病毒 PP31 原核表达 多克隆抗体 

分 类 号：S476.13[农业科学—农业昆虫与害虫防治]