华北农学报Issue(4)：90-94,5.DOI:10.7668/hbnxb.2015.04.016
类猪圆环病毒 P1 VP1蛋白在昆虫细胞中的表达及特性分析
Expression and Characterisation of Porcine Circovirus-like Virus P1 VP1 Protein in Insect Cells
摘要
Abstract
To identify whether the porcine circovirus-like virus P1 VP1 protein can self-assemble into virus-like particles(VLPs)in vitro,P1 ORF1 gene was amplified and cloned into the pFastBac TM1 vector.Then P1 ORF1 gene was expressed in Sf9 cells with liposome transfection method.The expression was identified on transcriptional and protein levels,respectively,using RT-PCR and Western Blot.The properties of P1 VP1 protein expressed here was observed using electron microscopy.The results showed that cytopathy on Sf9 cells appeared on the 5th day after transfection,the 362 bp target fragments was amplified by RT-PCR from total RNA extracted from the cell inocula-tion and treated by recombinant DNase I.And the results showed that the cell culture after 72 h of inoculation could work with monoclonal anti-P1 VP1 specific reaction with Western Blot,and the inclusion bodies oval-like formed by P1 VP1 protein in Sf9 cells was revealed with electron microscopy.Porcine circovirus-like P1 VP1 protein was suc-cessfully expressed with baculovirus expression vector system,and no VLPs were observed,which contributes to the further study of the immunological functions of P1 VP1 protein.关键词
P1 VP1/昆虫杆状病毒表达载体系统/RT-PCR/Western Blot/电镜Key words
P1 VP1/Baculovirus expression vector system/RT-PCR/Western Blot/SEM分类
农业科技引用本文复制引用
王凤芝,温立斌,何孔旺,姚火春,王小敏..类猪圆环病毒 P1 VP1蛋白在昆虫细胞中的表达及特性分析[J].华北农学报,2015,(4):90-94,5.基金项目
国家自然科学基金项目（31272574；30972184），江苏省农业科技自主创新项目（CX（14）2045） （）
