猪伪狂犬病毒湖北株gE基因的克隆与序列分析

目的]通过对从湖北省某猪场分离获得的伪狂犬病毒的gE基因克隆和测序,分析其遗传变化规律。方法]参考GenBank中猪伪狂犬病病毒(Porcine pseudorabies Virus,PRV)gE基因的序列设计1对引物,对PRV湖北株gE基因进行PCR扩增和序列分析,PCR产物克隆到pMD 18-T载体。对重组质粒进行限制性内切酶分析和基因测序,与GenBank收录的国内外其他地区的标准参考毒株进行同源性分析和比较。结果]与标准参考毒株的核苷酸序列同源性为95.0%~98.0%,进化树分析表明,PRVHBXS2014株与河南焦作株EU561349-China同属于一个分支,亲缘关系较近,但存在个别位点的基因突变。结论]为有效防控该病提供参考。

Cloning of Hubei Strain gE Gene of Porcine Pseudorabies Virus and Sequence Analysis

Objective ] Based on cloning and sequencing of Hubei strain gE gene of porcine pseudorabies virus isolated from a pig farms in Hubei Province, their genetic variation was analyzed. Method ] A pairs of primers were designed and the sequence of gE gene of pseudorabies virus in the reference GenBank, PCR amplification and sequence analysis of PRV Hubei strain gE gene was done, and the PCR product was cloned into pMD 18-T vector. By restriction enzyme analysis and sequencing of recombinant plasmid, homology analysis and comparison with the other regions included in GenBank standard reference strains at home and abroad. Result] The results showed that with the standard reference strains, the nu- cleotide sequence homology of 95.0% -98.0%, phylogenetic tree analysis showed that, PRVHBXS2014 strain and Jiaozuo EU561349-China in Henan strain belonged to the same branch, close relationship,but there are some gene mutation, and the results were consistent with the reported in the literature. Conclusion] The study provided a reference for effective prevention and control of the disease.