| 构建携带EGFP标志人白细胞介素10基因的重组腺病毒载体 |
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| 引用本文： | 林健清,林献忠,张志坚. 构建携带EGFP标志人白细胞介素10基因的重组腺病毒载体[J]. 临床麻醉学杂志, 2009, 25(1) |
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| 作者姓名： | 林健清 林献忠 张志坚 |
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| 作者单位： | 350005,福州市,福建医科大学附属第一医院麻醉科;350005,福州市,福建医科大学附属第一医院神经内科 |
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| 基金项目： | 福建省科技厅青年人才创新基金，福建省科技厅自然科学基金 |
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| 摘 要： | 目的 构建携带EGFP标志人白细胞介素10(hIL-10)基因腺病毒载体.方法 以pSNAV2.0-hIL-10重组质粒为模板.PCR扩增hIL-10基因,获得hIL-10 cDNA片段,并酶切连接到带有含强绿色荧光蛋白基因(EGFP)标记的pDC316-IRES-EGFP-lacZalpha载体质粒上,Pmel线性化重组质粒pDC316-hIL-10-IRES-EGFP,与腺病毒包装系统AdMax转染293包装细胞,包装产生复制缺陷型重组腺病毒,经反复感染293细胞扩增病毒后,进行离子交换法纯化病毒,并测定病毒颗粒数、滴度.结果 经PCR鉴定、限制性酶切分析及序列测定,证明已正确构建重组穿梭质粒pDC316-hIL-10-IRES-EGFP和重组腺病毒质粒Ad-hIL-10-IRES-EGFP.扩增纯化后.测得重组腺病毒颗粒数为3.2×1011VP/ml,OD260/OD280值约为2.0,滴度为1.1×1010TCID50/ml.结论 成功构建了重组腺病毒质粒Ad-hIL-10-IRES-EGFP,为进一步研究hIL-10基因奠定实验基础.
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| 关 键 词： | 白细胞介素10 腺病毒载体 |
Construction of the recombinant adenovirus vector carrying human interleukin-10 marked enhanced green fluourescent protein |
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| LIN Jian-qing,LIN Xian-zhong,ZHANG Zhi-jian. Construction of the recombinant adenovirus vector carrying human interleukin-10 marked enhanced green fluourescent protein[J]. The Journal of Clinical Anesthesiology, 2009, 25(1) |
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| Authors: | LIN Jian-qing LIN Xian-zhong ZHANG Zhi-jian |
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| Affiliation: | LIN Jian-qing,LIN Xian-zhong,ZHANG Zhi-jian.Department of Anesthesiology,First Affiliated Hospital,Fujian Medical University,Fuzhou 350005,China |
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| Abstract: | Objective To construct the recombinant adenovirus vector carrying human interleukin-10(hIL-10) marked enhanced green fluourescent protein(EGFP).Methods The hIL-10 gene was constructed by PCR with pSNAV 2.0-hIL-10 recombinant plasmid as template,enzyme digestion and ligation and inserted the obtained hIL-10 cDNA fragment into shuttle vector pDC316-IRES-EGFP-lacZalpha linearization with PmeI and supercoil adenovirus skeleton plasmid pAdMax for homologous recombination.The 293 cells were transfected with the o... |
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| Keywords: | Interleukin-10 Adenovirus vector |
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