牛泡沫病毒BFV3026细胞感染性的确立及包装细胞系的建立  

Characterization of BFV3026 Infect ion and Construction of the Packag ing Cell Line

作　　者：余荭[1] 孔晓红[1] 李汀[1] 马永钢[1] 陈启民[1] 耿运琪[1] 

机构地区：[1]南开大学生命科学院,天津300071

出　　处：《病毒学报》2003年第4期330-335,共6页Chinese Journal of Virology

基　　金：国家自然科学基金(39970033)

摘　　要：分别将牛泡沫病毒BFV3026接种于胎牛肺细胞(FBL)、牛肺细胞系(BL12)、新生牛肾细胞(NBK)、兔肺细胞(RL)、人乳腺癌上皮细胞(MCF)、293T、HeLa、CV-1、CHO等9种细胞,通过对它们及其传代细胞的病变观察与RT-PCR检测,确立BFV3026对这9种细胞的感染。并以BFV3026原病毒DNA为模板,通过PCR构建以pcD NA3 1(-)为载体的gag-pol、env真核表达质粒共转染BL12细胞,经G418持续筛选,获得8个NeoR细胞克隆。RT-PCR及包装实验证实:其中7个细胞克隆能有效行使包装辅助功能。Fetal bovine lung cell(FB L),bovine lung cell(BL12),new born bovine kidney cell (NBK),rabbit lu ng cell(RL),CV-1,MCF,293T,HeLa and CHO cells were infected with bovine foam y virus(BFV3026).Virus infection i n these 9 cell lines was confirmed by detection of the cytopathic eff ects(syncytium formation)and BFV internal promoter(IP)regi on by RT-PCR in the infected cells and their passaged cells.In addition,gag-pol and env genes were cloned from the full-length DNA of BFV3026 and use d to co-transfect BL12.Stable bovi ne packaging cell line had been ge nerated by subjecting transfectant s to neomycin selection and screen ing for coincident expression of t hree structure genes using RT-PCR. The capacity of package was measur ed with transfecting of BFV transf er vehicle into the packaging cell lines and infecting of the recover ed defective viruses into fresh B L12 cells,which contain luciferase report gene cloned under the contr ol of BFV IP.

关 键 词：牛泡沫病毒 BFV3026细胞 感染性 包装细胞系 

分 类 号：S852.659.3[农业科学—基础兽医学]