›› 2008, Vol. 1 ›› Issue (1): 39-43.

• 生物技术 • 上一篇    下一篇

两株高致病性PRRSV ORF6和ORF7基因克隆及序列分析

时建立1,2,李俊2,徐绍建1,2,赵鸿雁1,2, 张秀美2 , 王金宝2
  

  1. 1青岛农业大学,青岛 266109; 2山东省农科院畜牧兽医研究所,济南 250100
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2008-01-20 发布日期:2008-01-20

Clone and Sequence Analysis of Two Highly Pathogenic PRRSV ORF6 and ORF7 Genes

SHI Jianli1,2,LI Jun2, XU Shaojian1,2, ZHAO Hongyan1, 2, ZHANG Xiumei2, WANG Jinbao2
  

  1. 1. Qingdao Agricultural University, Qingdao 266109,China;2.Institute of Animal Science and Veterinary Medicine,Shandong Academy of Agricultural Sciences, Jinan 250100,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2008-01-20 Published:2008-01-20

摘要: 应用RTPCR方法从实验室分离的两株高致病性PRRSV SX、ZQ株中扩增出ORF6和ORF7,将其分别克隆、测序。用DNAStar 软件分析所测序列,并与VR2332株、LV株、周边国家及国内分离株进行核苷酸和推导氨基酸同源性比较,并绘制系统进化树,结果ORF6、ORF7核苷酸与北美洲型的同源性为91.1%~100%,与欧洲型的同源性为66.2%~70.7%,推导氨基酸与北美洲型的同源性为91.2%~100%,与欧洲型的同源性为62.3%~82.3%。证明新分离到的PRRSV SX株、ZQ株仍属北美洲型。SX株ORF6、ORF7核苷酸与国内新分离到的高致病性PRRSV JXA1株同源性分别为99.8%、100%;ZQ株ORF6、ORF7核苷酸与国内新分离到的高致病性PRRSV JXA1株同源性分别为99.6%、99.7%。

关键词: 高致病性PRRSV; ORF6; ORF7; 克隆; 序列分析

Abstract: The ORF6 and ORF7 genes of two highly pathogenic PRRSV SX,ZQ,derived from Shandong,were amplified by RT-PCR.The products were sequenced after cloning into pMD18-T vector respectively .The sequences were analysed using DNAStar and compared with those of VR-2332,LV and other strains isolated from China and circumjacent Country.The homologous of ORF6,ORF7 Nuclear acid was 91.1%~100% with North American(VR-2332) and 66.2%~70.7% with European(LV) viruses, The homologous of ORF6,ORF7 deduced amino acid was 91.2%~100% with North American(VR-2332) and 62.3%~82.3% with European(LV) viruses,indicating that the two new shandong isolates were related to the North American PRRSV genotype. The ORF6,ORF7 Nuclear acid of SX shared 99.8%,100% identity with highly pathogenic PRRSV JXA1,while The ORF6,ORF7 Nuclear acid of ZQ shared 99.6%,99.7% identity with highly pathogenic PRRSV JXA1.

Key words: highly pathogenic PRRSV; ORF6; ORF7; clone; sequence analysis

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