| 猪繁殖与呼吸综合征病毒SGQ株M蛋白基因截断片段原核表达载体构建及其表达的初步研究 |
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| 引用本文： | 陈进会,冯迎春,颜其贵,黄伟,韩国全,万莉,雷燕,王志彪.猪繁殖与呼吸综合征病毒SGQ株M蛋白基因截断片段原核表达载体构建及其表达的初步研究[J].动物检疫,2010(12):37-39,44. |
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| 作者姓名： | 陈进会 冯迎春 颜其贵 黄伟 韩国全 万莉 雷燕 王志彪 |
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| 作者单位： | [1]东莞出入境检验检疫局,广东东莞523072 [2]动物疫病与人类健康四川省重点实验室,四川农业大学,四川雅安625014 [3]四川农业大学动物医学院,四川雅安625014 |
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| 基金项目： | 基金项目：东莞市科技局项目（2007108101100） |
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| 摘 要： | 为表达猪繁殖与呼吸综合征病毒（PRRSV）M蛋白主要抗原表位基因序列，参照GenBank中发表的PRRSVSCQ株M蛋白基因设计并合成一对特异性引物，通过PCR方法从重组质粒pMD18-T-M扩增得到缺失N端跨膜区的M蛋白基因片段dM（deletingM），将其与pMD19-Tsimplevector连接，经测序正确后克隆至高效原核表达载体pGEX-4T-1，得到重组表达载体pGEX-4T-1-dM，并将其转化大肠杆菌Rosetta（DE3），经IPTG于37℃诱导，PRRSVM基因获得表达。经SDS-PAGE分析，所表达的融合蛋白分子量约为35kDa。以纯化的重组蛋白作为抗原，经WesternBlot分析结果表明该重组蛋白可被PRRSV阳性血清所识别，可用于PRRSV的检测。
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| 关 键 词： | 猪繁殖与呼吸综合征病毒 M蛋白基因 原核表达 |
The Construction of Prokaryotic Expression Vector of M Protein Gene Truncated Fragment of Porcine Reproductive and Respiratory Syndrome Virus SCQ Strains and Preliminary Studies for its Expression |
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| Authors: | Chen Jinhui Feng Yinchun Yan Qigui Huang Wei Han Guoquan Wan Li Lei Yan Wang Zhibiao |
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| Institution: | 1.Dongguan Entry-Exit Inspection and Quarantine Bureau, Dongguan, Guangdong,523072 China; 2.Sichuan Province Key Laboratory of Animal Disease and Human Health, Ya＇an, Sichuan, 625014 China; 3.College of Animal Medicine, Sichuan Agriculture university, Ya＇an, Sichuan, 625014 China) |
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| Abstract: | According to the PRRSV SCQ strain M gene nucleotide sequence reported in the GenBank, a pair of specific primers was designed and synthesized in order to successful expression of M protein of porcine reproductive and respiratory syndrome virus （PRRSV） ,the gene segment dM （deleting M）deleting N transmembrane domain sequence was successfully amplified from recombinant plasmids pMD18-T-M by PCR. Then, the dM gene was cloned into plasmid of PGEX-4T-1 and transformed into Rosetta （DE3）. PRRSV M gene was expressed by IPTG induction at 37℃. SDS analysis showed that molecular weight of recombinant protein was about 35 kDa. The recombinant protein was analyzed by Western-blot, the result demonstrated that the recombinant protein was recognized by the PRRSV positive serum, so the M protein could be used for the detection of PRRSV. |
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| Keywords: | PRRSV M protein gene prokaryotic expression |
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