表达乙脑病毒PrM基因重组伪狂犬病病毒的构建  

Construction of Recombinant Virus Strain TK^-/gG^-/PrM^+ Recombinized by Pseudorabies Virus Ea Mutant Strain TK^-/gG^-/LacZ^+ and PrM Gene of Japanese Encephalitis Virus

作　　者：李自力[1] 陈焕春[1] 徐高原[1] 方六荣[1] 肖少波[1] 王祥[2] 何启盖[1] 

机构地区：[1]华中农业大学动物医学院湖北省预防兽医学重点实验室,武汉430070 [2]湖北省公安厅,武汉430034

出　　处：《畜牧兽医学报》2007年第1期53-58,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：教育部重大项目(0113)

摘　　要：本研究以乙型脑炎病毒SA14-14-2疫苗株基因组RNA为模板,采用RT-PCR一步法扩增了PrM基因的全长cDNA(558 bp),用BamHI和EcoRI双酶切PrM基因的扩增产物,回收目的基因后将其克隆至经同样酶切的伪狂犬病病毒通用转移载体pPgG-uni中,获得了转移载体pPgG-PrM,并对其外源片段进行了测序。序列分析结果表明:与已报道的乙型脑炎病毒SA14强毒株和SA14-14-2疫苗株的核苷酸序列比较,PrM基因的同源性为100%。以伪狂犬病病毒Ea株TK-/gG-/LacZ+突变株为载体构建了一株表达乙型脑炎病毒PrM基因的重组伪狂犬病病毒TK-/gG-/PrM+,为进一步开展猪乙型脑炎与伪狂犬病二价基因工程疫苗的研究奠定了基础。Based on the nucleotide sequence of JEV SA14-14-2 strain, a pair of primers was designed. The PrM gene of JEV was cloned into general transfer vector pPgG-uni. The sequence analysis showed that the PrM genes of SA14 and SA14-14-2 strains had 100% homology. A cotransfection experiment was carried out with the purified pPgG-PrM and the genome of TK^-/ gG^-/LacZ^＋ mutant of pseudorabies virus Ea strain in PK-15 cells. By plaque purification and PCR detection, the recombinant virus TK^-/gG^-/PrM^＋ was purified. This recombinant virus strain can be used for the study of duel-valence vaccine of JEV and PRV.

关 键 词：乙型脑炎病毒 PrM基因 重组伪狂犬病病毒 

分 类 号：S852.65[农业科学—基础兽医学]