新城疫病毒F基因真核表达载体的构建及瞬时表达  

Construction and transient expression of Newcastle disease virus F gene Eukaryotic vector in COS-7 cell line

作　　者：王青[1] 张彦明[1] 王选年[2] 张改平[3] 胡建和[2] 杨苏珍[3] 赵东[3] 邢广旭[3] 徐彦召[1] 赵光辉[4] 段艳华[4] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100 [2]河南科技学院动物科学学院,河南新乡453003 [3]河南省农业科学院河南省动物免疫学重点实验室,河南郑州450002 [4]河南农业大学牧医工程学院,河南郑州450002

出　　处：《西北农林科技大学学报（自然科学版）》2007年第10期6-10,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家"863"计划项目(2003AA249031)

摘　　要：应用PCR扩增出新城疫病毒F基因,将其克隆入pGEM-T载体,测序。然后将NDVF基因与高效的真核表达载体pcDNA4/HisMax相连,构建NDVF基因重组真核表达质粒pc4F,采用Superfect转染试剂将pc4F瞬时转染COS-7细胞,应用细胞免疫组化法检测其在细胞中的表达情况。结果表明,F基因能够在COS-7细胞中成功表达。The F gene of NDV was amplified by PCR and cloned into pGEM-T Easy vector and sequenced. NDV F gene was, then,subcloned into high efficient eukaryotic vector pcDNA4/HisMax. The recombinant plasmid pc4F was transfected into COS-7 cells by using superfect transfection reagent. The NDV F protein expression in COS-7 cells was detected by immunocytochemistry. The transient expression of NDV-F protein provided the foundation for stable expression in a cell line,and for the future preparation of monoclonal antibodies that could be used to detect NDV and viral antigen variation. Results showed that F gene could be successflly experessed in COS-7 cells.

关 键 词：新城疫病毒 F基因 真核表达 

分 类 号：S858.31[农业科学—临床兽医学] S852.659.5[农业科学—兽医学]