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Expression vector construction and protocaryon expression of rotavirus VP4 gene and VP7 gene
Author(s): WANG Pengyan1, CHEN Chuangfu1, YU Xinglong2, XU Xingran2, TU Changchun2, ZHANG Gaoxuan1(1.animalscienceandtechnologycollege, SHIHEZI University, SHIHEZI 832003, china, 2. Themilitaryveterinaryinstitute, QUARTERMASTER Universityofpla, CHANGCHUN 130062, china)
Pages: 7-
12
Year: 2003
Issue:
1
Journal: Chinese Journal of Veterinary Science and Technolo
Keyword: VP4基因; VP7基因; 载体构建; 原核表达;
Abstract: 经PCR从含有VP4基因、VP7基因片段的重组质粒 pT VP4和 pT VP7中扩增VP4、VP7基因 ,连接至 pGEM 5zf(+ )。经筛选和鉴定正确后同时酶切目的基因和表达质粒 pET 2 8a(+ ) ,连接、转化受体菌 ,经PCR、酶切和序列分析证明 ,连接向位和阅读框架是正确的 ,从而构建了轮状病毒保护性抗原VP4基因、VP7基因片段的原核表达载体 pET2 8a VP4、pET2 8a VP7。重组质粒转化表达菌在IPTG诱导下 ,用SDS PAGE、薄层凝胶扫描分析表达的蛋白。结果表明 ,表达的目的蛋白以包涵体的形式存在 ,蛋白的分子量分别为 37.6 9和 36 .2 0ku ,表达量占菌体总蛋白的比例分别为 17.1%和 14 .4 %。
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