摘要
通过RT-PCR和重组PCR技术扩增出猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)疫苗株sM、M、S基因,构建重组腺病毒穿梭质粒。穿梭质粒与BJ5183细菌同源重组构建同源重组腺病毒质粒,同源重组腺病毒质粒经PacⅠ酶切后转染AD-293细胞,获得3个含有目的基因的重组腺病毒。将3个具有感染能力的复制缺陷性重组腺病毒共同感染Vero细胞,细胞上清液进行小鼠免疫特性研究。结果表明,共同感染Vero细胞所得蛋白能诱导小鼠产生特异性体液免疫应答。
sM,M,S gene of PEDV vaccine stain were implified by amplified by RT-PCR and reorganization PCR,then inserted into the pShuttle-CMV vector.These recombinant shuttle plasmids were linearized by PmeⅠand then transformed to BJ5183 cells containing the pAdEasy-1 bone plasmid.The homologous recombinantplasmidwas transformed to XL-10 Gold ultracompetent cells in order to obtain this plasmid in a large scale.These recombinant adenoviruses were co-infected Vero cells,mice immune experiment indicated that antibodies were induced by Western blotting.
出处
《中国畜牧兽医》
CAS
北大核心
2012年第2期11-16,共6页
China Animal Husbandry & Veterinary Medicine
基金
江西省教育厅2010年度科技项目(GJJ10602)
