扬州大学学报（农业与生命科学版）2017，Vol.38Issue(3)：6-10,5.DOI:10.16872/j.cnki.1671-4652.2017.03.002
表达羊口疮病毒B2L基因重组腺病毒的构建及鉴定
Construction and identification of recombinant adenovirus expressing B2L gene of the orf virus
摘要
Abstract
In order to construct a recombinant adenovirus expressing B2L gene of orf virus (orf virus,ORFV),the DNA was extracted from the ORFV AH-F10 clinical isolates.The B2L gene was amplified by PCR and then subcloned into the adenovirus shuttle vector plasmid.The recombinant shuttle plasmid was named pHBAd-ORFV-B2L.And it was eon-transfeeted with the backbone vector pHBAd-BHGloxAE1 to HEK293 cells.Forty hours post the transfection,the recombinant adenovirus was successfully packaged and named rAd-ORFV-B2L.Then,the viral titers and genetic stability of rAd-ORFV-B2L were tested.The results demonstrated that the titer of the rAd-ORFV-B2L was 1079TCID50 · μL-1.Western-blot assay suggested that rAd-ORFV-B2L could successfully express B2L proteins in vitro.The expressed B2L protein could react with the ORFV polyclonal antibody.The results of RT-PCR from series passages showed that rAdORFV-B2L could stably express the exogenous B2L protein.This study laid the foundation for the animal protection experiment using recombinant adenovirus in the future.关键词
羊口疮病毒/B2L基因/重组腺病毒/构建/鉴定Key words
ORFV/B2L gene/recombinant adenovirus/construction/identification分类
农业科技引用本文复制引用
王勇,蒋书东,李永东,杨侃侃,王元红,俞赵荣,潘飞,崔佣楸,苏莉,钟灵毓,徐前明..表达羊口疮病毒B2L基因重组腺病毒的构建及鉴定[J].扬州大学学报（农业与生命科学版）,2017,38(3):6-10,5.基金项目
国家自然科学基金资助项目(31602063) （31602063）
安徽省自然科学基金资助项目(1508085QC60) （1508085QC60）
安徽农业大学稳定和引进人才科研项目(YJ2015-16) （YJ2015-16）
