中国兽医科学2017，Vol.47Issue(11)：1385-1391,7.DOI:10.16656/j.issn.1673-4696.2017.11.007
猪圆环病毒2d亚型病毒样颗粒的制备与鉴定
Preparation and identification of porcine circovirus type 2d Cap protein as virus-like particles in insect cells
摘要
Abstract
The capsid protein(Cap) comprises the major structural protein of porcine circovirus type 2(PCV2).In this study,a full-length of Cap gene was cloned and sequenced for phylogenic analysis of PCV2 strain and antigenicity analysis of Cap protein,and then Cap gene was expressed via Bac to Bac expression system in insect cells.The target protein was identified by Western blot,IFA,MS,EM and Immuno-EM methods in following work.lt was shown that the cloned Cap gene was classified into a PCV2d subtype gene cluster as the reference strains through phylogenic analysis.By antigenicity analysis,four domains were found to be different with those from domestic PCV2a,PCV2b and PCV2 vaccine strains.By the Bac to bac expression system,PCV2d Cap protein was highly expressed in insect cells.The target protein was identified with anti-Cap monoclonal antibody by WBand analyzed by MS method.Antigenicity was analyzed with PCV2 positive serum through IFA assay.Finally,the target protein was checked as virus-like particles Via electron microscopy and immuno-electron microscopy.All together,PCV2d VLP was successfully prepared and identified in insect cells and had good immunogenicity as a PCV2d sub-unit vaccine candidate or diagnostic reagent in future work.关键词
猪圆环病毒2d亚型/核衣壳蛋白/杆状病毒表达系统/病毒样颗粒Key words
porcine circovirus type 2/capsid protein/baculovirus expression system/virus-like particle分类
农业科技引用本文复制引用
侯强,侯绍华,贾红,姜一瞳,鑫婷,李明,陈青,朱鸿飞..猪圆环病毒2d亚型病毒样颗粒的制备与鉴定[J].中国兽医科学,2017,47(11):1385-1391,7.基金项目
国家重点发展计划项目(2017YFD0502302) （2017YFD0502302）
北京市教育委员会科技计划面上项目(KM201410020002) （KM201410020002）
中国农业科学院科技创新工程(ASTIP-IAS-11) （ASTIP-IAS-11）
