柯萨奇病毒B3型3D蛋白抗体的制备  

Preparation of coxsackievirus B3 3D protein polyclonal antibody

作　　者：王玉[1] 徐维祯[1] 穆宇松 王天楹 闫碧莹[1] 颜学勤[1] 赵文然[2] 钟照华[1] 

机构地区：[1]哈尔滨医科大学微生物学教研室及伍连德研究所,哈尔滨150081 [2]哈尔滨医科大学细胞生物学教研室,哈尔滨150081

出　　处：《微生物与感染》2017年第3期172-178,共7页Journal of Microbes and Infections

基　　金：国家自然科学基金(81571999;81672007;31300144)

摘　　要：肠道病毒3D蛋白是其RNA聚合酶。柯萨奇病毒B3型(coxsackievirus B3,CVB3)主要感染心脏,其3D蛋白在心肌表达中的时序和分布尚不清楚。本研究将通过聚合酶链反应(polymerase chain reaction,PCR)获得的CVB 3D片段插入pET28a(+)的表达框,获得pET28a(+)-3D重组质粒。异丙基β-D-硫代半乳糖苷(isopropylβ-D-1-thiogalactopyranoside,IPTG)诱导pET28a(+)-3D表达3D-His蛋白,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate-polyacrylamide gel electrophoresis,SDS-PAGE)后,切胶,获得3D-His蛋白。3D-His蛋白加佐剂免疫新西兰大白兔制备3D蛋白多克隆抗体,蛋白免疫印迹法检测抗体效价及特异性。结果显示,本研究获得了高效价且特异性好的抗CVB3 3D蛋白抗体,可用于CVB3 3D蛋白功能的后续研究。Coxsackievirus B3 （CVB3） mainly infects the heart, and the timing and distribution of viral RNA polymerase （3D protein） expression in the myocardium are currently unclear. In this study, the CVB3 3D gene was obtained by polymerase chain reaction （PCR） and cloned into the prokaryotic expression vector pET28a（ ＋ ）. The expression of 3D was induced by isopropyl [3-D-l-thiogalactopyranoside （IPTG）, and the 3D-His protein was purified and used to immunize New Zealand white rabbits for antiserum against the 3D protein. An anti-CVB3 3D antibody with high titer and specificity was obtained, and it can be used for future functional study of CVB3 3D protein.

关 键 词：柯萨奇病毒B3型 3D聚合酶 抗体 原核表达 

分 类 号：R392[医药卫生—免疫学]