Expression of the M and N Protein Gene of Porcine Reproductive and Respiratory Syndrome Virus in E ．coli

CA Jia-Li; CA Bao-Xiang; JIANG Beng

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June 1999

CA Jia-Li, CA Bao-Xiang and JIANG Beng. Expression of the M and N Protein Gene of Porcine Reproductive and Respiratory Syndrome Virus in E ．coli[J]. Virologica Sinica, 1999, 14(3): 244-248.

Citation: CA Jia-Li, CA Bao-Xiang, JIANG Beng. Expression of the M and N Protein Gene of Porcine Reproductive and Respiratory Syndrome Virus in E ．coli .VIROLOGICA SINICA, 1999, 14(3) : 244-248.

猪繁殖和呼吸综合征病毒膜蛋白和核衣壳蛋白基因在大肠杆菌中的表达

1.
南京农业大学动物医学院，南京210095
2.
西南农业大学动物养殖学院，重庆400716

摘要

以ＰＲＲＳＶ弱毒株膜蛋白（Ｍ）和核衣壳（Ｎ）蛋白基因为模板，设计的一对含有ＥｃｏＲＩ和ＢａｍＨＩ酶切位点的引物，通过ＲＴＰＣＲ扩增出一约９００ｂｐ的ＭＮ基因片段，将此基因片段成功克隆于高效表达载体ｐＢＶ２２０，构建成重组质粒ｐＢＶＭＮ，导入大肠杆菌ＤＨ５，经温敏诱导，成功地表达了ＭＮ基因。表达产物经ＳＤＳＰＡＧＥ电泳和Ｗｅｓｔｅｒｎｂｌｏｔ印迹分析，其分子量约３４ｋＤ，与兔抗ＰＲＲＳＶ高免血清发生反应，经光密度扫描分析，表达产物量占菌体总蛋白的１２％。该研究为ＰＲＲＳ基因诊断抗原的研制奠定了基础
- 猪繁殖和呼吸综合征病毒
- , 质粒pBV220
- , 表达
- , M蛋白
- , N蛋白

Expression of the M and N Protein Gene of Porcine Reproductive and Respiratory Syndrome Virus in E ．coli

1.
Animal Medical College，N iang Agricultural University，Nanjang 210095

Abstract

The primers for RT PCR were designed on the basis of M and N gene sequence of PRRSV. A gene fragment about 900 bp, which has digestion sites of EcoRI and BamHI, was amplified by RT PCR. The M and N gene were cloned into expression vector pBV220 and one recombinant PBVMN was constructed which highly expressed a 34 kD protein in \%E. coli\% DH 5 . The expressed product was identified by SDS PAGE and Western blotting, which occupies 12% of total bacterial protein. The report has laid a basis for the development of molecular diagnostic antigen of PRRSV.
- Porcine reproduetive and respiratory syndrome virus(PRRSV)
- , pBV220
- , Expres sion
- , M protein
- , N protein

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Expression of the M and N Protein Gene of Porcine Reproductive and Respiratory Syndrome Virus in E ．coli

1. Animal Medical College，N iang Agricultural University，Nanjang 210095

Abstract: The primers for RT PCR were designed on the basis of M and N gene sequence of PRRSV. A gene fragment about 900 bp, which has digestion sites of EcoRI and BamHI, was amplified by RT PCR. The M and N gene were cloned into expression vector pBV220 and one recombinant PBVMN was constructed which highly expressed a 34 kD protein in \%E. coli\% DH 5 . The expressed product was identified by SDS PAGE and Western blotting, which occupies 12% of total bacterial protein. The report has laid a basis for the development of molecular diagnostic antigen of PRRSV.