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| IL-7和EGFP双基因共表达重组腺病毒载体的构建 | |
| 引用本文： | 宁昌,余长林,李建军,胡锴勋. IL-7和EGFP双基因共表达重组腺病毒载体的构建[J]. 现代肿瘤医学, 2011, 19(12): 2401-2404. DOI: 10.3969/j.issn.1672-4992.2011.12.12 |
| 作者姓名： | 宁昌 余长林 李建军 胡锴勋 |
| 作者单位： | 1. 解放军161医院肿瘤科,湖北,武汉,430010 2. 解放军军事医学科学院附属307医院血液科,北京,100071 3. 解放军总政治部门诊部,北京,100011 |
| 摘 要： | 目的:构建白细胞介素7(IL-7)和增强型绿色荧光蛋白(EGFP)共表达的重组腺病毒载体,为进一步感染干细胞奠定基础。方法:将目的基因IL-7克隆到含有报告基因EGFP的穿梭质粒中,然后再将构建的重组穿梭质粒转移至pAdxsi载体中,构建重组腺病毒载体质粒,继而在H293细胞中扩增,纯化后测定病毒滴度。结果:pShuttle-EGFP-mIL7重组穿梭质粒经酶切鉴定得到0.5kb和5.1kb 2条带;pAdxsi-EGFP-mIL7重组腺病毒载体质粒经酶切鉴定得到14K、11.8K、3.1kb、2.66kb、2.47K、1.45K、0.6K 7条带;TCID50法测定纯化后的病毒滴度为2×1010pfu/ml。结论:pAdxsi-EGFP-mIL7重组腺病毒载体构建成功。 |
| 关 键 词： | 白细胞介素7 增强型绿色荧光蛋白 重组腺病毒载体 构建 |
Construction of recombinant adenoviral vector co-expressing interleukin-7 and enhanced green fluorescent protein |
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| NING Chang,YU Changlin,LI Jianjun,HU Kaixun. Construction of recombinant adenoviral vector co-expressing interleukin-7 and enhanced green fluorescent protein[J]. Journal of Modern Oncology, 2011, 19(12): 2401-2404. DOI: 10.3969/j.issn.1672-4992.2011.12.12 | |
| Authors: | NING Chang YU Changlin LI Jianjun HU Kaixun |
| Affiliation: | NING Chang1,YU Changlin 2,LI Jianjun3,HU Kaixun2 1People's Liberation Army 161 Hospital Oncology,Hubei Wuhan 430010,China,2 Department of Heamatologe,Affiliated Hospital of Academy of Military Medical Science,Beijing 100071,3 Department of Clinics,People's Liberation Army Gerneral Political,Beijing 100011 |
| Abstract: | Objective:To construct the adenoviral vector co-expressing interleukin-7(IL-7) and enhanced green fluorescent protein(EGFP),to lay a experimental foundation for further study on the infection into stem cell.Methods: The target gene IL-7 was cloned into the shuttle plasmid expressed the report gene EGFP.Then the recombinant shuttle plasmid was transformed into Dh5a bacteria to recombine with backbone vector pAdxsi.Next,the plasmid pAd-EGFP-mIL7 was amplified in H293 cells and purifired,then the viral titer w... |
| Keywords: | IL-7 EGFP recombinant adenoviral vector construction |
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