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.
出处
《生物技术通报》
CAS
CSCD
北大核心
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)
关键词
小菜蛾颗粒体病毒
PP31
原核表达
多克隆抗体
PluteUa xylostella granulovirus （ PlxyGV ） PP31
Prokaryotic expression
Polyclonal antibody
