羊痘病毒属病毒ORF132的克隆及序列分析

目的为得到实验室现有的山羊痘、绵羊痘、疙瘩皮肤病病毒的ORF132基因序列,以便于后续研究。方法本实验在参考GenBank公布的羊痘病毒、疙瘩皮肤病病毒ORF132基因序列的基础上,设计了3对引物,对1株山羊痘病毒,1株绵羊痘病毒,两株疙瘩皮肤病病毒进行PCR扩增,克隆扩增产物,进行测序分析。结果测序结果在NCBI网站进行比对后发现其与标准毒株有较大相似性,同时存在不同的变异现象。结论羊痘病毒属的3种病毒同源性极高,相比之下,ORF132基因具有较好的特异性,通过该次的基因分析,可为PCR引物设计和基因芯片探针设计提供基础。

Cloning and sequence analysis of capripoxvirus ORF132

In order to get the ORF132 gene sequences of goat pox,sheep pox and lump skin disease viruses in our laboratory to facilitate follow-up study.These three strains of viruses were proliferated in goat testicular cells.Then three pairs of primers were designed referring the published ORF132 gene sequences of these three viruses in GenBank.The DNA of a strain of goat pox,a strain of sheep pox virus and two strains of lump skin disease viruses were extracted and amplified via PCR.At last,the amplified products were cloned and sequenced and the sequencing results were compared in the NCBI website.In this analyzed cases,the cloning goatpox virus was 95% similar with goatpox virus strain G20-LKV and Pellor strain,and the sheeppox was 98% similar to sheeppox virus A strain and NISKHI strain;lump skin disease vaccine strain was 99% similar to isolate Neethling vaccine LW 1959 strain;lump skin disease wild strain was 99% similar to the NI-2490 isolate Neethling 2490 strain or NW-LW isolate Neethling Warmbaths LW strain.Viruses belonging to capripoxvirus are highly homologous to each other,however,ORF132 genes are with high specificity.This genetic analysis provides the basis for PCR primer design and microarray probe design.