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猪生殖与呼吸综合征病毒 M基因和N 基因的表达
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赖维莉，郭万柱，颜其贵，韩国全，欧 洋
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（四川农业大学 动物生物技术中心，四川 雅安 625014）
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摘要：
将猪生殖与呼吸综合征病毒（PRRSV）的M基因和N基因从重组质粒pMD18-T-M-N中亚克隆至pBV220原核表达载体上，成功构建了重组表达质粒pBVM-N。将pBVM-N转化大肠埃希氏菌DH5 α 感受态细胞，重组菌经温度敏感诱导表达，其细菌裂解物经SDS-PAGE可检测到分子质量约为19 ku的目的蛋白，与M基因表达产物一致;经蛋白质分析软件Bandscan分析，其表达量可达19.1%;Western-blotting分析表明，该重组蛋白可被兔抗PRRSV血清所识别，与预期的M基因表达产物相一致，而N基因未获表达。 |
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关键词：
猪生殖与呼吸综合征病毒；M基因；N基因；原核表达 |
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中图分类号：S 852.65: Q 786文献标识码：
A 文章编号：1000-6419(2005)10-0771-04 |
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Prokaryotic expression of M and N genes of porcine reproductive and respiratory syndrome virus |
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LAI Wei-li，GUO Wan-zhu，YAN Qi-gui，HAN Guo-quan，OU Yang |
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（Center of Animal Biotechniques，Sichuan Agricultural University，Ya'an 625014，China） |
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Abstract: After the membrane protein gene（ORF6） and nucleocapsid protein gene（ORF7） in the recombinant plasmid pMD18-T-M-N were subcloned into a prokaryotic expression vector pBV220，the re-combinant plasmid pBVM-N was constructed successfully. The pBVM-N was transformed into E.coli DH5 α and the recombinant E.coli DH5 α cells were induced by temperature. The SDS-PAGE test indicated that the membrane protein gene was expressed successfully in a high level and the expressed non-fusion protein was about 19 ku in molecular weight. The Western-blotting test showed that the protein had immunologically reactive activity，which was in accordance with the expected result. However the nucleo-capsid protein gene was not expressed. |
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Key words: porcine reproductive and respiratory syndrome virus（PRRSV）；membrane protein gene（ORF6）；nucleocapsid protein gene（ORF7）；prokaryotic expression |
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