伪狂犬病病毒gN基因缺失株的构建及其在小鼠上的免疫效力分析  

Construction and Immune Response of Pseudorabies Virus Mutants with gN Gene Deletion

作　　者：胡艳芬[1] 管萌[1] 刘开春[1] 陈素娟[1] 彭大新[1] 刘秀梵[1] 

机构地区：[1]扬州大学兽医学院农业部畜禽传染病学重点开放实验室禽类预防医学教育部重点实验室,扬州225009

出　　处：《畜牧兽医学报》2014年第1期87-93,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：江苏省青蓝工程项目(苏教师2010[27]号);教育部“长江学者和创新团队发展计划”创新团队项目(IRT0978);江苏高校优势学科建设工程

摘　　要：为了确定伪狂犬病病毒(PRV)gN基因的功能,以PRV-JSZ双基因缺失株rPRV-gE^-/TK^-和疫苗株Bartha K61为亲本株,通过同源重组技术构建这两种病毒的gN缺失株。PCR扩增鉴定和序列分析结果显示成功获得了gN缺失株rPRV-gE^-/TK^-/gN^-和rPRV-Ba-gN^-。与亲本株相比,rPRV-gE^-/TK^-/gN^-在PK15细胞上早期增殖速度较慢,但效价无明显变化;rPRV-Ba-gN^-在PK15细胞上的增殖速度和效价显著降低。rPRVgE^-/TK^-/gN^-和rPRV-gE^-/TK^-的LD_50均大于1×10~6 TCID_50,rPRV-Ba-gN^-的毒力低于Bartha K61。与亲本株相比,gN缺失株免疫小鼠可提供较好的抗体水平和攻毒保护率。PRV gN基因的缺失可进一步降低病毒的毒力,提高免疫保护力。The aim of this study was to determine the role of gN gene of pseudorabies virus(PRV).The PRVs rPRV-gE^-/TK^- and Bartha K61 were selected as parental strains,and mutants with gN gene deletion were constructed by homologous recombination.PCR amplification and sequence analysis showed that the mutants with gN gene deletion(rPRV-gE^-/TK^-/gN^-and rPRV-Ba-gN^-) were constructed successfully.Compared with parental strain,growth speed of rPRV-gE^-/TK^- /gN^- was slightly slow in the early stage and its titer was not affected on PK15 cells,while both growth speed and titer of rPRV-Ba-gN^- were significantly lower on PK 15cells.The LD_(50) of both strains rPRV-gE^-/TK^- /gN^- and rPRV-gE^- /TK^- were more than 1×10~6TCID_(50),the virulence of rPRV-Ba-gN^- was attenuated compared with the parental strain Bartha K61.The two gN deletion mutants could provide higher antibody level and protection against virulent PRV challenge than the parental strains.The deletion of gN gene in PRV may contribute to decreased virulence and increased immune protection.

关 键 词：伪狂犬病病毒 gN基因 毒力 免疫保护 

分 类 号：S852.659.1[农业科学—基础兽医学]