| 表达胰高血糖素样肽1重组腺伴随病毒载体的构建 |
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| 引用本文： | 娄明武,梁冰,张明东,杨广笑,王全颖.表达胰高血糖素样肽1重组腺伴随病毒载体的构建[J].中国组织工程研究与临床康复,2011,15(28). |
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| 作者姓名： | 娄明武 梁冰 张明东 杨广笑 王全颖 |
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| 作者单位： | 1. 深圳市龙岗中心医院介入影像科,广东省深圳市,518116
2. 西安市华广生物工程有限公司,陕西省西安市,710025 |
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| 基金项目： | 广东省科学技术厅资助项目，深圳市科技和信息局资助项目，深圳市龙岗区委组织部2008年专家提升计划资助项目 |
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| 摘 要： | 背景:胰高血糖素样肽1(glucagon-like peptide-1,GLP-1)在人体内半衰期过短限制了其应用.目的:构建可表达GLP-1的重组腺伴随病毒.方法:将NT4-GLP-1融合基因插入腺伴随病毒包装质粒pSSHG-CMV中,构建pSSHG/NT4-GLP-1重组腺伴随病毒包装质粒.采用磷酸钙共沉淀法将辅助质粒pAAV/Ad、腺病毒质粒pFG140及pSSHG/NT4-GLP-1转染至293细胞系,用其感染Hela细胞.结果与结论:重组质粒pSSHG/NT4-GLP-1经限制性内切酶EcoRⅠ酶切鉴定可见342 bp的目的片段,说明NT4-GLP-1融合基因已经成功重组于腺伴随病毒包装质粒pSSHG-CMV内.免疫细胞化学结果显示,转染pSSHG/NT4-GLP-1重组腺伴随病毒包装质粒的Hela细胞内有大量棕黄色颗粒,阳性率达到70%以上,说明NT4-GLP-1重组腺伴随病毒在细胞中可以表达GLP-1.
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| 关 键 词： | 胰高血糖素样肽1 腺伴随病毒 重组质粒 构建 组织构建 |
Construction of a recombinant adeno-associated virus plasmid expressing glucagon-like peptide-1 |
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| Lou Ming-wu,Liang Bing,Zhang Ming-dong,Yang Guang-xiao,Wang Quan-ying.Construction of a recombinant adeno-associated virus plasmid expressing glucagon-like peptide-1[J].Journal of Clinical Rehabilitative Tissue Engineering Research,2011,15(28). |
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| Authors: | Lou Ming-wu Liang Bing Zhang Ming-dong Yang Guang-xiao Wang Quan-ying |
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| Abstract: | BACKGROUND:Glucagon-like peptide-1 (GLP-1) is limited by half-life in human body. OBJECTIVE:To construct a recombinant adeno-associated virus vector expressing GLP-1. METHODS:The fusion gene NT4-GLP-1 was inserted into the adeno-associated virus packaging plasmid pSSHG-CMV to generate the adeno-associated virus packaging plasmid pSSHG/NT4-GLP-1. 80% of fused 293 cells were co-transfected by using the calcium phosphate precipitation method, with aid packaging plasmids pAAV/Ad, pFG140 and the recombinant aden... |
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