| A型口蹄疫病毒VP0基因的克隆及原核表达 |
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| 引用本文： | 刘明,刘航,柳纪省.A型口蹄疫病毒VP0基因的克隆及原核表达[J].生物技术通报,2012(1):104-107. |
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| 作者姓名： | 刘明 刘航 柳纪省 |
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| 作者单位： | 中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部兽医公共卫生重点开放实验室,兰州,730046 |
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| 摘 要： | 从实验室冻存的含A型口蹄疫病毒的细胞中提取FMDV总RNA,通过RT-PCR获得cDNA.并根据FMDV全基因组序列设计了一对针对VP0基因的引物,通过PCR扩增得到目的基因VP0并亚克隆入pMD18-T载体.将鉴定出的阳性质粒和表达载体pET32a用BamH Ⅰ和HindⅢ双酶切回收后连接获得阳性重组质粒pET32-VP0.用IPTG诱导重组质粒表达目的蛋白VP0并用SDS-PAGE进行检测.表达产物用镍亲和树脂进行了纯化.结果证明,口蹄疫病毒VP0蛋白在大肠杆菌中获得了高效表达且表达产物得到了纯化,为实验室进一步的研究提供了重要的材料.
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| 关 键 词： | 口蹄疫病毒 VP0基因 原核表达 |
Cloning and Prokaryotic Expression of VP0 Gene of Foot-and-mouth Disease Virus (FMDV) Type A |
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| Liu Ming
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Liu Hang
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Liu Jixing.Cloning and Prokaryotic Expression of VP0 Gene of Foot-and-mouth Disease Virus (FMDV) Type A[J].Biotechnology Bulletin,2012(1):104-107. |
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| Authors: | Liu Ming Liu Hang Liu Jixing |
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| Institution: | Liu Ming Liu Hang Liu Jixing(Key Laboratory of Veterinary Public Health of the Ministry of Agriculture,State Key Laboratory of Veterinary Etiological Biology,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046) |
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| Abstract: | Extract the RNA of FMDV tape A from frozen cells was extracted,and then the cDNA was acquired by RT-PCR.A special primer pair was designed which contain BamHⅠand HindⅢ according to complete genome of FMDV.The target gene VP0 was amplified by PCR and subcloned into pMD18-T vector.pMD-VP0 and expression vector pET-32a were digested by with BamHⅠ and HindⅢ;gene VP0 was ligated into pET-32a,which formed recombinant plasmid pET32-VP0 after identification.The interest gene was induced to express in E.coli with IPTG and identified with SDS-PAGE.The target protein was purified with Ni-NTA Purification System.Result showed that the target protein was expressed in E.coli and also purified. |
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| Keywords: | Foot-and-mouth disease virus VP0 gene Prokaryotic expression |
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