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表达猪繁殖与呼吸综合征病毒GP5蛋白重组伪狂犬病毒的构建及其生物学特性分析
引用本文:田志军,仇华吉,倪健强,周艳君,蔡雪辉,周国辉,王云峰,童光志.表达猪繁殖与呼吸综合征病毒GP5蛋白重组伪狂犬病毒的构建及其生物学特性分析[J].遗传学报,2005,32(12):1248-1255.
作者姓名:田志军  仇华吉  倪健强  周艳君  蔡雪辉  周国辉  王云峰  童光志
作者单位:中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,哈尔滨,150001
基金项目:国家高技术研究发展计划项目(“863”计划)(编号:2001AA213051)和国家重点基础研究发展规划项目(“973”计划)(编号G1999011902)资助
摘    要:将猪繁殖与呼吸综合征病毒(Porcine Reproductive and Respiratory Syndrome Virus,PRRSV)CH-1a株GP5基因插入到伪狂犬病病毒(Pseudorabies Virus,PRV)Bartha-K61株TK基因中,获得了一株TK^-/gE^-表型的重组伪狂犬病病毒,命名为rPRV—GP5。经生长动力学、表达动力学和间接免疫荧光证实PRRSV GP5在重组病毒中获得了表达,表达蛋白的抗原性与亲本病毒相似,rPRV-GP5在不同的细胞上毒价和细胞病变与Bartha—K61比较无显著差异。4只PRV抗体阴性的绵羊,每只接种10^6.0。PFU的rPRV-GP5可以完全抵抗10^3LD50伪狂犬病强毒S株的攻击。10头PRV、PRRSV抗体阴性的仔猪滴鼻接种rPRV-GP5 10^7.0 PFU/头并在接种后63d攻击PRRSV CH-1a株10^5.0 TCID50/头,攻毒后3d和5d出现了PRRSV荧光抗体和ELISA抗体,在政毒后14d检测到了PRRSV中和抗体,Bartha—K61活疫苗组和对照组至实验结束时仍未检测到PRRSV中和抗体。这说明rPRV-GP5免疫产生了针对PRRSV的回忆性免疫应答。

关 键 词:猪呼吸与繁殖综合征病毒  伪狂犬病病毒  GP5  生物学特性
文章编号:0379-4172(2005)12-1248-08
收稿时间:2005-03-15
修稿时间:2005-03-152005-06-08

Construction and Characterization of a Recombinant Pseudorabies Virus Expressing Porcine Reproductive and Respiratory Syndrome Virus GP5
TIAN Zhi-Jun,QIU Hua-Ji,NI Jian-Qiang,ZHOU Yan-Jun,CAI Xue-Hui,ZHOU Guo-Hui,WANG Yun-Feng,TONG Guang-Zhi.Construction and Characterization of a Recombinant Pseudorabies Virus Expressing Porcine Reproductive and Respiratory Syndrome Virus GP5[J].Journal of Genetics and Genomics,2005,32(12):1248-1255.
Authors:TIAN Zhi-Jun  QIU Hua-Ji  NI Jian-Qiang  ZHOU Yan-Jun  CAI Xue-Hui  ZHOU Guo-Hui  WANG Yun-Feng  TONG Guang-Zhi
Institution:National Key Laboratory of Veterinary Biotechnology, Harbin Veterinar Research Institute, CAAS, Harbin 150001, China
Abstract:The GP5 gene of porcine reproductive and respiratory syndrome virus (PRRSV) was integrated into the TK gene locus of pseudorabies virus (PRV) vaccine strain Bartha-K61,resulting in a TK- and gE- negative recombinant PRV harboring GP5 gene,designated as rPRV-GP5.The in vitro expression of the GP5 by rPRV-GP5-infected cells was analyzed by single-step growth analysis,Western blot,and indirect immunofluorescence test.It was shown that GP5 gene can be expressed authentically in the cytoplasm of rPRV-GP5-infected cells.Compared to its parental virus,rPRV-GP5 showed no obvious difference regarding viral replication and cytopathogenic effects in several cell cultures.Four PRV-negative sheep immunized intramuscularly with 106.0 PFU of rPRV-GP5 were fully protected from challenge with 103 LD50 of highly virulent PRV S strain of porcine origin.Ten PRV- and PRRSV-negative piglets given intranasally with 107.0 PFU of rPRV-GP5 and challenged intranasally with 105.0 TCID50 of virulent PRRSV CH-1a strain at day 63 post-inoculation developed antibodies against PRRSV 3,5,14 days post-challenge,as revealed by indirect immunofluorescence test,enzyme-linked immunosorbent assay and virus neutralization test.The results suggest that rPRV-GP5 is capable of inducing anamnestic immune response to PRRS in inoculated animals.
Keywords:porcine reproductive and respiratory syndrome virus (PRRSV)  pseudorabies virus (PRV)  GP5  characterization
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