| 猪传染性胃肠炎病毒N蛋白基因的克隆及其原核表达载体的构建 |
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| 引用本文： | 翁崇鹏,张莉,毛娅卿,何后军.猪传染性胃肠炎病毒N蛋白基因的克隆及其原核表达载体的构建[J].江西农业大学学报,2004,26(4):576-580. |
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| 作者姓名： | 翁崇鹏 张莉 毛娅卿 何后军 |
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| 作者单位： | 1. 江西农业大学动物科学技术学院,江西南昌,330045
2. 北京市农科院畜牧兽医研究所,北京,100089 |
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| 基金项目： | 国家"863"资助项目(2003AA241121002) |
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| 摘 要： | 以本实验室从河北分离到的TGEV毒株,采用RT-PCR技术扩增了TGEV N基因,长为1149bp,并进行亚克隆重组到pMD18-T质粒载体上,连接,转化,鉴定后得到阳性重组质粒．命名为pTN．将重组质粒插入的片段进行序列分析和比较,结果表明TGEV河北分离株N基因与国外的Purdue-115、FS772/70、TO14、96-1933株均具有较高的同源性,达95％以上。推导的氨基酸序列同源性为95％以上。并将目的基因与原核表达载体pGEX-4T-3重组,成功构建了原核表达载体pGEX-N。pGEX-N表达载体的构建,为其蛋白质的表达提供重要依据。
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| 关 键 词： | 猪传染性胃肠炎病毒 N基因 分子克隆 载体构建 |
| 文章编号： | 1000-2286(2004)04-0576-05 |
| 修稿时间： | 2004年4月25日 |
Cloning and Sequencing of N Gene from Transmissible Gastroenteritis Virus and the Constructed Prokaryotic Expressing Plasmid |
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| WENG Chong-peng,ZHANG Li,MAO Ya-qing,He Hou-jun.Cloning and Sequencing of N Gene from Transmissible Gastroenteritis Virus and the Constructed Prokaryotic Expressing Plasmid[J].Acta Agriculturae Universitis Jiangxiensis,2004,26(4):576-580. |
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| Authors: | WENG Chong-peng ZHANG Li MAO Ya-qing He Hou-jun |
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| Institution: | WENG Chong-peng~1,ZHANG Li~2,MAO Ya-qing~1,He Hou-jun~1 |
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| Abstract: | The viral subgenome mRNA of porcine transmissible gastroenteristis virus(TGEV) isolated from Hebei was amplified with RT-PCR. A DNA fragment was amplified which contains completely ORF of N gene, and the size of the DNA fragment is about 1 149bp.The PCR products were purified and then cloned into plasmid pMD18-T cloning vector, the recombinant plasmids were designated pTN and analyzed by endonecleoase digestion from proper inserts. The sequence analysis of the insert fragment in the recombinant pTN indicated that TGEV isolated from Hebei shared more than 95% with Purdue-115 strain, FS772/70 strain, TO14 strain,96-1933 strain. The amino acid sequence homology was above 95%. The N gene was subcloned into pGEX-4T-3, an prokaryotic expressing vector. Molecular cloning of N gene from TGEV provides a basis for the studies on protein expressing and molecular characteristic and the constructed expressing plasmid pGEX-N can be used in futher protein expressing. |
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| Keywords: | TGEV N gene molecular cloning prokaryotic expressing plasmid |
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