中国兽医科学2017，Vol.47Issue(3)：315-321,7.DOI:10.16656/j.issn.1673-4696.2017.03.007
脑心肌炎病毒VP1基因重组腺病毒的构建及对小鼠免疫效果的研究
Construction of recombinant adenovirus with encephalomyocarditis virus VP1 gene and its immune effect on mice
摘要
Abstract
By construction of recombinant adenovirus with encephalomyocarditis virus(EMCV) VP1 gene,the study aims to offer experimental basis for EMCVvaccine development.EMCV VP1 gene was amplified,and linked with pYr-adshuttle-4 to construct the shuttle vector pYr-ads-4-VP1.The shuttle vector with adenovirus vector by in vitro recombination was constructed,designed as recombinant adenovirus vector pAd-VP1-EGFP,which was finally packaged into HEK293 cells successfully to obtain recombinant adenovirus rAd-VP1-EGFP.Cells were infected with this recombinant adenovirus,then VP1 in the mRNA and protein levels.Finally,mice were immunized with rAd-VP1-EGFP,and its immune effects were verified.The results showed that the recombinant adenovirus rAd-VP1-EGFP was successful,and TCID50 was 10-8.5/mL.VP1 was expressed in gene both the mRNA and protein levels after infected with recombinant adenovirus.The survival rate indicated that rAd-VP1-EGFP had a good immune effect on mice,significantly higher than single immunination.The above-results laid the foundation for further EMCV genetic engineering vaccine development.关键词
脑心肌炎病毒/结构蛋白VP1/腺病毒/免疫效果Key words
encephalomyocarditis virus(EMCV)/structural protein VP1/adenovirus/immune effect分类
农业科技引用本文复制引用
张海霞,马忠仁,冯若飞,令瑛,杨术伟,凡静静,李琼毅,李向茸,马玉梅,包晓婧,魏嘉..脑心肌炎病毒VP1基因重组腺病毒的构建及对小鼠免疫效果的研究[J].中国兽医科学,2017,47(3):315-321,7.基金项目
国家自然科学基金项目(31460665) （31460665）
教育部“长江学者和创新团队发展计划”项目(IRT13091) （IRT13091）
中央高校基本科研业务费专项资金项目(31920150030,3192015005,ZyP2015005) （31920150030,3192015005,ZyP2015005）
