| 人GDF-5重组腺病毒载体的构建及其在人间充质干细胞中的表达 |
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| 引用本文： | 程相俊,刘浩,薛建新,张霆. 人GDF-5重组腺病毒载体的构建及其在人间充质干细胞中的表达[J]. 生物医学工程学杂志, 2010, 0(1) |
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| 作者姓名： | 程相俊 刘浩 薛建新 张霆 |
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| 作者单位： | 四川大学华西医院骨科;四川大学华西医院肿瘤科;四川大学生物治疗国家重点实验室; |
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| 基金项目： | 国家自然科学基金资助项目(30572163) |
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| 摘 要： | 本研究目的在于构建携带GDF-5基因的重组腺病毒表达载体,并用其感染人骨髓来源的间充质干细胞(MSCs)研究GDF-5的表达。从含有人GDF-5基因核心序列的质粒pCMV-SPORT6上通过PCR扩增GDF-5,将其酶切连接到穿梭质粒pAdtrack-CMV上,在BJ5183中和骨架质粒pAdeasy-1同源重组,筛选阳性克隆并酶切鉴定,线性化后磷酸钙法转染入人胚肾293细胞中包装、扩增,得到含有GDF-5基因的重组腺病毒并测定其滴度。用收集的腺病毒感染靶细胞MSCs,在基因水平检测GDF-5的表达情况。结果表明,成功构建了含有GDF-5基因的重组腺病毒载体,病毒滴度为1×109PFU/ml。重组腺病毒能有效感染MSCs并表达目的基因。通过构建该腺病毒并感染人MSCs可得到能持续一定时间表达GDF-5蛋白的MSCs,为这种转基因的MSCs进一步修复组织奠定了基础。
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| 关 键 词： | 生长分化因子5 间充质干细胞 重组腺病毒 基因治疗 |
Construction of Human GDF-5 on Adenovirus Vector and Its Expression in MSCs |
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| Cheng Xiangjun Liu Hao Xue Jianxin Zhang Ting. Construction of Human GDF-5 on Adenovirus Vector and Its Expression in MSCs[J]. Journal of biomedical engineering, 2010, 0(1) |
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| Authors: | Cheng Xiangjun Liu Hao Xue Jianxin Zhang Ting |
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| Affiliation: | Cheng Xiangjun1 Liu Hao1 Xue Jianxin2 Zhang Ting31(Department of Orthopaedics,West China Hospital of Sichuan University,Chengdu 610041,China)2(Department of Cancer,China)3(State Key Laboratory of Biotherapy of Sichuan University,China) |
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| Abstract: | This experimental study was aimed to construct the recombinant adenovirus vector containing human GDF-5 gene,and to use it for infecting human MSCs and detecting the expression of the gene GDF-5.The core sequence of human GDF-5 was amplified by PCR from pCMV-SPORT6,and then was cloned to pAdtrack-CMV.The linearized shuttle plasmid pAdtrack-CMV-GDF-5 was homogenously recombined with pAdeasy-1 in BJ5183.The potential clone was analyzed by restriction endonuclease digestion.The correct clone was linearized and... |
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| Keywords: | Growth and differentiation factors-5(GDF-5) Mesenchymal stem cells(MSCs) Recombinant adenovirus Gene therapy |
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