猪瘟病毒E^(rns)基因的克隆与序列分析

Cloning and Sequence Analysis of Erns Gene of CSFV

为了解山东地区猪瘟病毒的变异情况,将6份经RT-PCR方法鉴定为猪瘟病毒(CSFV)阳性的山东地区患病猪的病料接种PK-15细胞,分离得到6株猪瘟病毒。对该6株猪瘟病毒的Erns基因主要抗原编码区序列进行了RT-PCR扩增、克隆、测序,并经DNA Star软件对测序结果与HCLV疫苗株、Shimen株及Alfort株Erns基因进行比较和分析,构建了CSFV遗传进化树。结果扩增的6株山东CSFV分离株Erns基因同源性为98.5%~100%,与HCLV疫苗株,Shimen株及Aflort株的同源性为82.9%~85.2%;6株CSFV分离株Erns蛋白的同源性为99.1%~100%,与HCLV疫苗株,Shimen株及Aflort株Erns蛋白的同源性分别为88.0%~88.4%和89.4%~89.8%。表明所分离的6株山东CSFV流行毒株的Erns基因发生了较大变异。

In order to get the message of the epidemiology and sequence variation of classical swine fever virus（CSFV）,six samples which were collected and detected by reverse transcriptase polymerase chain reaction（RT-PCR） from Shandong province were found to be positive to CSFV.The six samples were inoculated in PK-15 cells for virus isolation,and six strains of CSFV were isolated.The major antigen coding region sequences of Erns gene of the six isolated CSFV were amplified by RT-PCR,then cloned and sequenced.The above-mentioned sequences were analyzed and compared with the same gene sequence of HCLV vaccine,Shimen and Alfort strains by DNA Star.The constructed phylogenetic tree showed that nucleotide homogeneity of Erns gene of six CSFV Shandong strains was 98.5%-100.0%.Nucleotide homogeneity of Erns gene of six CSFV Shandong strains with Erns gene of HCLV vaccine,Shimen and Alfort strains ranged from 82.9%-85.2%.Amino acid homology of Erns gene of six CSFV Shandong strains was 99.1%-100%,while 88.0%-88.4% with HCLV vaccine strain and 89.4%-89.8% with Shimen and Alfort strains.The results suggested that mutation occurred in Erns gene of the six CSFV Shandong strains.