| 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NDV F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。 |
| 关键词: 新城疫病毒 F基因 真核表达 |
| DOI： |
| 分类号: |
| 基金项目:国家“863”计划项目(2003AA249031) |
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| Construction and transient expression of Newcastle disease virus F gene Eukaryotic vector in COS-7 cell line |
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| Abstract: |
| 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. |
| Key words: NDV F gene eukaryotic expression |
