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| 河北猪繁殖与呼吸综合征病毒HB-3(cz)株N基因变异分析及原核表达 | |
| 引用本文： | 陈赛娟,刘涛,曹洪战,张斌,付新城,张丽青,孙继国.河北猪繁殖与呼吸综合征病毒HB-3(cz)株N基因变异分析及原核表达[J].河北农业大学学报,2008,31(5). |
| 作者姓名： | 陈赛娟 刘涛 曹洪战 张斌 付新城 张丽青 孙继国 |
| 作者单位： | 1. 河北农业大学,动物科技学院,河北,保定,071001 2. 商务部流通产业促进中心,北京,100062 3. 廊坊市畜牧水产局,河北,廊坊,102800 |
| 摘 要： | 为了给河北地区预防和控制猪繁殖与呼吸综合征提供理论数据。应用RT-PCR方法特异性扩增HB-3(cz)株的ORF7(N)基因片段,将扩增片段克隆入pMD-T载体后测序,应用DNAstar分析软件对序列进行分析,并与GenBank中发表的PRRSV毒株序列进行比较。结果显示:HB-3(cz)株与高热病毒株JXA1、HUB2等及传统河北分离株HB-1(sh)氨基酸同源性高达97.6%,属美洲型毒株。将目的片段克隆入原核表达载体pGEX-6P-1,重组质粒pGEX-N转化大肠杆菌BL21(DE3),在IPTG诱导下成功获得表达,经Western-blot-ting分析表明:表达的融合蛋白分子量约为39.5 kDa,能与PRRSV的阳性血清发生特异性反应,为PRRSV血清学诊断方法的建立奠定了基础。 |
| 关 键 词： | 猪繁殖与呼吸综合征病毒 N基因 序列分析 原核表达 |
Sequence analysis and prokaryotic expression of ORF7 gene of porcine reproductive and respiratory syndrome virus HB-3(cz) strain |
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| CHEN Sai-juan,LIU Tao,CAO Hong-zhan,ZHANG Bin,FU Xin-cheng,ZHANG Li-qing,SUN Ji-guo.Sequence analysis and prokaryotic expression of ORF7 gene of porcine reproductive and respiratory syndrome virus HB-3(cz) strain[J].Journal of Agricultural University of Hebei,2008,31(5). | |
| Authors: | CHEN Sai-juan LIU Tao CAO Hong-zhan ZHANG Bin FU Xin-cheng ZHANG Li-qing SUN Ji-guo |
| Abstract: | This study aims to provide theoretical evidence for prevention and control of porcine reproductive and respiratory syndrome in Hebei.The specific ORF7 gene of HB-3(cz) strain amplified by RT-PCR was cloned into pMD-T vector.The recombinant plasmid was sequenced and compared to the other published strains in GeneBank.The results showed the deduced amino acids of HB-3(cz) showed 97.6% similarity to strains JXA1,HUB2,HB-1(sh),and so on,and the HB-3(cz) strain be1ongs to North American genotype.The ORF7 gene was cloned into prokaryotic expression vector pGEX-6P-1.The recombinant fusion proteins pGEX-N were highly expressed in E.coli BL21 in the forms of inclusion bodies induced by IPTG at 28℃.The expressed protein was identified to be 39.5 ku by SDS-PAGE analysis.Western-blotting analysis indicated that the recombinant protein can vigorously react against the positive serum of PRRSV,which lays the foundation for the development of the diagnosis methods in serology for PRRSV. |
| Keywords: | porcine reproductive and respiratory syndrome virus ORF7 gene sequence analysis prokaryotic expression |
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