| 小鼠脂联素与绿色荧光蛋白融合基因表达载体的构建及在COS-7细胞中的表达 |
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| 引用本文： | 王会中,付秋霞,高明,贾帅争,詹林盛,王全立.小鼠脂联素与绿色荧光蛋白融合基因表达载体的构建及在COS-7细胞中的表达[J].生物技术通讯,2005,16(5):492-495. |
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| 作者姓名： | 王会中 付秋霞 高明 贾帅争 詹林盛 王全立 |
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| 作者单位： | 1. 军事医学科学院,输血医学研究所,北京,100850;解放军第305医院,北京,100017
2. 军事医学科学院,输血医学研究所,北京,100850 |
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| 摘 要： | 以绿色荧光蛋白(GFP)基因(gfp)为报告基因,构建小鼠脂联素(mADPN)基因(mAd)与gfp的融合基因mAd/gfp表达载体pCI-neo-apoEHCR-hAATp-mAd-gfp,脂质体法转染体外培养的COS-7细胞,荧光显微镜观察GFP在细胞中的表达可间接反映mADPN的表达,并通过RT-PCR在核酸水平进一步确证mAd的表达.荧光显微镜观察及RT-PCR结果均证明mADPN在COS-7细胞中获得了高效表达,表明mADPN重组表达载体pCI-neo-apoEHCR-hAATp-mAd可以在真核细胞COS-7中高效表达mADPN,为进一步探讨mAd在小鼠体内的表达提供了可行性依据.
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| 关 键 词： | 脂联素 绿色荧光蛋白 融合基因 真核表达 |
| 文章编号： | 1009-0002(2005)05-0492-04 |
| 收稿时间： | 03 14 2005 12:00AM |
| 修稿时间： | 2005年3月14日 |
Construction of Eukaryotic Expression Vector of Mouse Adiponectin/GFP Fusion Gene and its Expression in COS-7 Cells |
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| WANG Hui-zhong,FU qiu-xia,GAO Ming,JIA Shuai-zheng,ZHAN Lin-sheng,WANG Quan-li.Construction of Eukaryotic Expression Vector of Mouse Adiponectin/GFP Fusion Gene and its Expression in COS-7 Cells[J].Letters in Biotechnology,2005,16(5):492-495. |
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| Authors: | WANG Hui-zhong FU qiu-xia GAO Ming JIA Shuai-zheng ZHAN Lin-sheng WANG Quan-li |
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| Institution: | 1.Institute of Transfusion Medicine. Academy of Military Medical Sciences, Beijing 100850; 2. The 305 Hospital of PLA, Beijing 100017, China |
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| Abstract: | To verify the expression of the mouse adiponectin(mADPN) gene(mAd) recombinant expression vector pCI-neo-apoEHCR-hAATp-mAd in mammalian cells. As a reporter, the green fluorescent protein(GFP) gene(gfp) was fused to the downstream of mAd, and the fusion gene mAd/gfp was cloned into the expressive vector pCI-neo-apoEHCR-hAATp to form the recombinant vector pCI-neo-apoEHCR-hAATp-mAd-gfp, which was transferred into COS-7 cells by liposome. The expression of GFP in COS-7 cells was observed by fluorescent microscopy, and the mRNA expression of mAd was detected by RT-PCR. Results showed that the recombinant mADPN was efficiently and stably expressed in COS-7 by vector pCI-neo-apoEHCR-hAATp-mAd-gfp. The recombinant vector pCI-neo-apoEHCR-hAATp-mAd can express mADPN in mammalian cells, and it's proper to be used in mouse liver. |
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| Keywords: | adiponectin green fluorescent protein fusion gene eukaryotic expression |
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