| 腭裂相关基因TTF-2转基因小鼠载体的构建及体外表达 |
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| 引用本文： | 黄磊,石冰,郑谦,王龑,宋庆高.腭裂相关基因TTF-2转基因小鼠载体的构建及体外表达[J].昆明医学院学报,2011,32(7):12-16. |
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| 作者姓名： | 黄磊 石冰 郑谦 王龑 宋庆高 |
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| 作者单位： | 1. 广州市第一人民医院口腔科,广州广东,510016
2. 四川大学华西口腔医学院唇腭裂研究室,四川成都,610041 |
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| 基金项目： | 国家自然科学基金资助项目(30171020) |
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| 摘 要： | 目的扩增及克隆C57BL/6j小鼠titf-2基因并利用小鼠骨髓间充质细胞进行表达.方法从C57BL/6j小鼠肝脏组织中提取基因组DNA,用PCR从中扩增出titf-2基因片段.将titf-2基因片段插入载体pMD18-T中,经全自动序列分析仪测序正确后,再克隆至表达载体pBROAD3-mcs中,构建重组表达载体pBROAD3-mcs-titf-2,并转染小鼠骨髓间充质细胞进行表达.用羊抗鼠TTF-2多抗IgG对表达产物进行Western Blot签定.结果 DNA测序结果证实,获得titf-2基因片段,大小与理论值相符;Western Blot分析表明,表达出TTF-2约为4.2KDa大小的蛋白.结论成功扩增、克隆titf-2基因,并在小鼠骨髓间充质细胞中得到表达,为下一步建立titf-2转基因小鼠模型奠定了基础.
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| 关 键 词： | TTF-2 克隆 Western Bloting |
Construction and in Vitro Expression of Cleft Palate Relevant Gene TTF-2 Transgenic Mice Vector |
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| HUANG Lei,SHI Bing,ZHENG Qian,WANG Yan,SONG Qing-gao.Construction and in Vitro Expression of Cleft Palate Relevant Gene TTF-2 Transgenic Mice Vector[J].Journal of Kunming Medical College,2011,32(7):12-16. |
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| Authors: | HUANG Lei SHI Bing ZHENG Qian WANG Yan SONG Qing-gao |
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| Institution: | HUANG Lei,SHI Bing,ZHENG Qian,WANG Yan,SONG Qing-gao(1)Dept.of Stomotology,The First People's Hospital of Guangzhou,Guangzhou Guangdong 510016,2)Cheilopalatognathus Research Room,West China College of Stomatology,Sichuan University,Chengdu Sichuan 610041,China) |
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| Abstract: | Objective To amplify and clone the C57BL/6J mouse titf-2 gene and express its protein in the mouse marrow mesenchymal cell.Methods The DNA encoding the C57BL/6J mouse titf-2 gene was amplified by PCR from the mouse liver tissue.The titf-2 gene was inserted into plasmid pMD18-T.After sequencing,The titf-2 gene was cloned into eukaryotic expression vector pBROAD3-mcs to construct the recombinant expression vector pBROAD3-titf-2 and it was used to transfect the mouse marrow mesenchymal cell.The expression prod... |
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| Keywords: | TTF-2 Cloning Western bloting |
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