华南农业大学学报2018，Vol.39Issue(2)：9-15,7.DOI:10.7671/j.issn.1001-411X.2018.02.002
猪伪狂犬病病毒流行毒株gE/gI缺失突变株的构建及生物学特性研究
Construction of agE/gI-deleted mutant strain of epidemic porcine pseudorabies virus and its biological characteristics
摘要
Abstract
[Objective]To obtain a candidate vaccine strain against epidemic porcine pseudorabies virus.[Method]A gE/gI-deleted transferring plasmid pMD-LA-RA and a recombinant plasmid carrying EGFP gene were constructed according to the sequence of epidemic porcine pseudorabies virus. The homologous recombination was operated between pMD-LA-EGFP-RA and PRV AH, and then the recombinant mutant virus PRV AHgE–/gI–/EGFP+ was selected using EGFP as screening marker. In order to obtain the gE/gI-deleted mutant strain PRV AHgE–/gI– without theEGFP gene, the second homologous recombination was carried out between p MD-LA-RA and PRV AH gE–/gI–/EGFP+. The proliferation ability, genetic stability and immunogenicity of PRV AH gE–/gI– were evaluated by its growth curve, continuous passage in susceptible cells and animal immunization.[Result]PRV AH gE–/gI– was obtained through two homologous recombinations with fluorescence observation, plaque purification and PCR. The deletion of gE,gI genes and EGFP gene marker in PRV AH gE–/gI– was identified by PCR, fluorescence observation and sequencing. PRV AH gE–/gI–had similar proliferation ability to the parental strain and had good genetic stability and immunogenicity.[Conclusion]A gE/gI-deleted mutant strain of epidemic porcine pseudorabies virus with good immunogenicity was constructed successfully, which provides a basis for the development of gene deleted vaccine targeting epidemic strains.关键词
猪伪狂犬病病毒/流行毒株/基因缺失/同源重组/生物学特性Key words
porcine pseudorabies virus/epidemic strain/gene deletion/homologous recombination/biological characteristic分类
农业科技引用本文复制引用
潘慧,李艳华,向柯宇,唐栋,程珍珠,罗永文,琚春梅..猪伪狂犬病病毒流行毒株gE/gI缺失突变株的构建及生物学特性研究[J].华南农业大学学报,2018,39(2):9-15,7.基金项目
广东省科技计划项目(2016A020210046,2016B090921009,2014A020214006,2015A020209100) （2016A020210046,2016B090921009,2014A020214006,2015A020209100）
