口蹄疫病毒VP1基因的原核可溶性表达  

The Exploration of Prokaryotic Soluble Expression of Foot and Mouth Disease Virus VP1 Gene

作　　者：王海烽[1,2] 易忠[1] 魏玉荣[1] 魏婕[1,2] 胡尔马西[1] 符子华[1] 

机构地区：[1]新疆畜牧科学院兽医研究所,乌鲁木齐830000 [2]新疆农业大学动物医学学院,乌鲁木齐830052

出　　处：《中国畜牧兽医》2009年第10期50-54,共5页China Animal Husbandry & Veterinary Medicine

摘　　要：针对重组原核表达载体表达目的蛋白以包涵体形式存在的问题,作者对影响外源蛋白表达的IPTG的浓度、温度、时间等因素均进行了探索,以观察VP1蛋白的可溶性情况。通过PCR技术将VP1基因克隆至pGEM-T载体中,然后将pET-28a(+)和pGEM-T载体分别进行双酶切,构建重组的pET-28a-VP1载体,将重组载体转化至BL21中,诱导后经SDS-PAGE检测可见约29 ku的目的蛋白条带。插入的外源目的蛋白主要以包涵体的形式存在,上清中的可溶性蛋白甚微。试验结果表明,这可能与蛋白本身的氨基酸组成有重要关联。Owing to the recombinant prokaryotic expression vector to express the target protein as inclusion body, VP1 protein was induced by different concentrations of IPTG,different temperatures and different time,cell lysates were analyzed by SDS-PAGE, to observe whether emerge soluble protein. The VP1 gene was amplified by PCR and cloned into pGEM-T vector, then pET-28a（＋ ） and pGEM-T vector were dual Enzyme digested respectively, a recombinant of pET-28a VP1 vector was a chieved and transformed into BL21. The results showed that protein expression yield was impacted by IPTG concentration and temperature and time, but the protein mainly was formed of inclusion bodies. The results show that inclusion body may be related to the amino acid composition.

关 键 词：口蹄疫病毒 VP1基因 原核 可溶性表达 

分 类 号：S852.659.6[农业科学—基础兽医学]