摘要
目的原核表达刚地弓形虫过氧化物氧化还原酶(peroxiredoxin,Prx)并制备多克隆抗体。方法 PCR技术扩增弓形虫cDNA中的prx基因,克隆至pET-28a(+)载体,构建prx/pET-28a(+)重组表达载体,转化至大肠埃希菌(E.coli)Rosetta中诱导表达。亲和层析纯化重组Prx蛋白,并制备兔多克隆抗体,蛋白印迹技术对多克隆抗体进行鉴定。结果成功从弓形虫cDNA中扩增出prx目的基因,构建了prx/pET-28a(+)重组质粒,获得抗Prx重组蛋白的多克隆抗体。蛋白印迹技术检测出弓形虫Prx的特异性条带。结论重组弓形虫Prx制备的多克隆抗体能检测Prx在弓形虫速殖子表达。
To clone and express the peroxiredoxin（Prx）of Toxoplasma gondii RH strain and produce the polyclonal antibody,prxgene fragments amplified by PCR were cloned into the pET-28a（＋）vector to construct the prokaryotic expression vector prx/pET-28a（＋）,which was efficiently expressed in E.coli Rosetta.Recombinant Prx proteins were purified by NiNTA affinity chromatography.The anti-Prx polyclonal antibody was produced in rabbit.Western blotting could detect Prx expression by anti-Prx polyclonal antibody.The recombinant Prx polyclonal antibody will facilitate the study of the biological functions of Prx.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2015年第12期1089-1092,共4页
Chinese Journal of Zoonoses
基金
国家自然科学基金(No.81301453)
卫生部寄生虫病原与媒介生物学重点实验室开放课题(No.WSBKTKT201302)
中国博士后科学基金特别资助(No.2015T80518)
中国博士后科学基金(No.2014M561598)
江苏省博士后科研计划(No.1402171C)
江苏大学高级人才启动基金(No.13JDG023
No.13JDG127)联合资助~~
