犬瘟热病毒GZ2株的分离鉴定及H基因原核表达质粒构建

对疑似CD的贵宾犬病料进行了CDV的分离和初步鉴定以及H基因的克隆和序列分析,以探讨不同分离株之间H基因的差异。结果表明:采用同步接种的方法,分离到1株犬瘟热病毒(CDV GZ2),并以其RNA为模板,扩增克隆了该病毒血凝素(H)基因,进而构建原核表达质粒pET-H。该毒株H基因ORF大小为1 824bp,可编码607个氨基酸;与国内分离株氨基酸一致性为97.4%～98.5%,与国外分离株为89.6%～95.6%,与疫苗株在89.6%～91.6%。进化分析表明:CDV的分布具有一定地域性,CDV GZ2与国内东北地区分离株亲缘关系近,而与国外分离株亲缘关系相对较远。

Isolation and Identification of the Canine Distemper Virus Guizhou Strain (CDV GZ2) and Prokaryotic Expression Plasmid Construction of the Viral Hemagglutinin Gene

The canine distemper virus(CDV GZ2) was isolated from suspicious CD pathologic material(poodle) and preliminarily identified,then the viral H gene was cloned and the sequence was analyzed to explore the difference of H gene among different isolated strains.The results showed that a CDV GZ2 strain was isolated by synchronization inoculation,and the viral H gene was cloned taking the viral RNA as templates,thus the prokaryotic expression plasmid was constructed.The viral H gene ORF was 1 824 bp,encoding 607 amino acids,97.4%～98.5% consistent with the strains in China,but 89.6%～95.6% with the strains abroad,and 89.6%～91.6% with vaccine strain.CDV GZ2 and the strains isolated from northeast area in China had the same ancestor,but the genetic relationships between CDV GZ2 and the strains from abroad were far.