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.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
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)
