| 人白细胞介素12双亚基共表达pVAX1-IRES-hIL12载体的构建与表达 |
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| 引用本文： | 王伟,高江平,阎瑾琦,肖毅,任杰,王宇,江乐,于继云.人白细胞介素12双亚基共表达pVAX1-IRES-hIL12载体的构建与表达[J].压电与声光,2010(6). |
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| 作者姓名： | 王伟 高江平 阎瑾琦 肖毅 任杰 王宇 江乐 于继云 |
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| 作者单位： | 军事医学科学院基础医学研究所;解放军总医院泌尿外科; |
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| 基金项目： | 国家“863”计划项目(2007AA02Z451);;国家自然科学基金项目(30772002,30840094)~~ |
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| 摘 要： | 目的构建下游可以共表达人白细胞介素12(hIL12)双亚基的双顺反子真核表达载体pVAX1-IRES-hIL12。方法通过搭桥PCR获得人白细胞介素12P35及P40双亚基的融合基因P35-F2A-P40,插入DNA疫苗载体pVAX1-IRES的下游,瞬时转染293-T细胞,ELISA检测融合基因的表达。结果酶切鉴定和序列分析表明融合基因与设计完全一致,融合基因在体外细胞培养液检测中获得分泌表达。结论该载体的成功构建可以为肿瘤基因疫苗研制提供免疫增效载体。
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| 关 键 词： | 白细胞介素12 基因融合 真核细胞 IRES |
Construction of human interleukin 12 double subunits co-expressing pVAX1-IRES-hIL12 vector and its expression |
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| WANG Wei,GAO Jiang-ping,YAN Jin-qi,XIAO Yi,REN Jie,WANG Yu,JIANG Le,YU Ji-yun Institute of Basic Medical Sciences,Academy of Military Medical Sciences,Beijing,China.Construction of human interleukin 12 double subunits co-expressing pVAX1-IRES-hIL12 vector and its expression[J].Piezoelectrics & Acoustooptics,2010(6). |
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| Authors: | WANG Wei GAO Jiang-ping YAN Jin-qi XIAO Yi REN Jie WANG Yu JIANG Le YU Ji-yun Institute of Basic Medical Sciences Academy of Military Medical Sciences Beijing China |
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| Institution: | WANG Wei1,2,GAO Jiang-ping2,YAN Jin-qi1,XIAO Yi1,REN Jie2,WANG Yu1,JIANG Le1,YU Ji-yun1 1 Institute of Basic Medical Sciences,Academy of Military Medical Sciences,Beijing 100850,China,2Department of Urology,Chinese PLA General Hospital,Beijing 100853 |
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| Abstract: | Objective To construct the bicistronic eucaryotic expression vector of human interleukin 12(hIL12)pVAX1-IRES-hIL12 co-expressing the hIL12 double subunits.Methods The fusion gene P35-F2A-P40,amplified by overlap extension PCR,was inserted into the downstream of the eukaryotic expression vector pVAX1-IRES.Then the recombinant plasmid pVAX1-IRES-hIL12 was transfected to 293T cells,and its expression was detected by ELISA.Results Enzyme digestion and sequence analysis showed that the bicistronic eucaryotic exp... |
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| Keywords: | hIL12 Gene Fusion Eukaryotic Cells IRES |
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