| 轮状病毒VP4基因和VP7基因原核表达载体的构建及表达 |
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| 引用本文： | 王鹏雁,陈创夫,余兴龙,徐兴然,涂长春,张高轩.轮状病毒VP4基因和VP7基因原核表达载体的构建及表达[J].中国兽医科学,2003,33(1):5-10. |
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| 作者姓名： | 王鹏雁 陈创夫 余兴龙 徐兴然 涂长春 张高轩 |
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| 作者单位： | 1. 石河子大学,动物科技学院,新疆,石河子,832003
2. 解放军军需大学,军事兽医研究所,吉林,长春,130062 |
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| 摘 要： | 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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| 关 键 词： | VP4基因 VP7基因 载体构建 原核表达 |
| 文章编号： | 1000-6419(2003)01-0005-06 |
| 修稿时间： | 2002年8月5日 |
Expression vector construction and protocaryon expression of rotavirus VP4 gene and VP7 gene |
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| WANG Peng yan ,CHEN Chuang fu ,YU Xing long ,XU Xing ran ,TU Chang chun ,ZHANG Gao xuan.Expression vector construction and protocaryon expression of rotavirus VP4 gene and VP7 gene[J].Veterinary Science in China,2003,33(1):5-10. |
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| Authors: | WANG Peng yan CHEN Chuang fu YU Xing long XU Xing ran TU Chang chun ZHANG Gao xuan |
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| Institution: | WANG Peng yan 1,CHEN Chuang fu 1,YU Xing long 2,XU Xing ran 2,TU Chang chun 2,ZHANG Gao xuan 1 |
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| Abstract: | The VP4 gene and VP7 gene were amplified from recombinant plasmids pT VP4 and pT VP7 by PCR and ligated with plasmid pGEM 5zf(+).The recombinant plasmids and expression vector pET 28a(+) were digested by restriction endonuclease.The two genes were ligated and transformed into E.coli.By PCR,RE digestion and sequencing the orientation and coding frame were correct.The result showed that the recombinant plasmids pET28a VP4?pET28a VP7were constructed. The transformed recombinant plasmids were induced by IPTG and the expressed proteins were valued by SDS PAGE with expected molecular size of 37.69 ku and 36.20 ku and the expression efficiency were respectively 17.1% and 14.4%. |
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| Keywords: | VP4 gene VP7 gene vector construction protocaryon |
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