猪瘟病毒兔化弱毒株与内蒙古分离野毒株(NMW)E2(gp55)基因的克隆与序列分析

Cloning and sequencing of E2(gp55)gene of prevalent virulent strain in Inner Mongolia autonomous region and C-strain of hog cholera virus

采用反转录PCR(RT-PCR)和套式PCR(nested-PCR)扩增了猪瘟病毒兔化弱毒株与内蒙古野毒株的E2(gp55)基因,片段长约1200bp,将PCR产物与PMD-18-T载体相连接并克隆后,经酶切、PCR鉴定后进行序列测定和分析,结果显示二者的核苷酸同源性为89.7%,这一结果表明内蒙古近期流行的猪瘟野毒株与目前使用的疫苗株在E2基因上存在一定的差异。

The major protective antigen E2 （gp55） gene of the C-strain strain of recent prevalent field virus in the Inner Mongolia autonomous transcription and nested polymerase chain reaction , both the amplified fragme （Chinese hog cholera virus ）and the region were amplified by reverse nts of E2 gene were about 1 200 bp in length by agarose gel electrophoresis. The two E2 fragments were cloned into PMD-18-T vector. They were identified by enzyme cleavage and PCR. Then the two kinds of E2 fragments were sequenced. The results of sequence analysis and comparison showed that the homology between the two E2 gene was 89.7 %. This result indicated that there were some difference in E2 gene between cell adapted Chinese hog cholera virus and the strain of recent prevalent field virus in the Inner Mongolia autonomous region.