| 真核表达载体pEGFP-C3-MASH-1的构建 |
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| 引用本文： | 赵杰,张瑞,许予明,赵国强,郭建新,宋波,韩志强. 真核表达载体pEGFP-C3-MASH-1的构建[J]. 医药论坛杂志, 2006, 27(8): 1-3,5 |
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| 作者姓名： | 赵杰 张瑞 许予明 赵国强 郭建新 宋波 韩志强 |
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| 作者单位： | 1. 郑州大学第一附属医院,郑州市,450052
2. 郑州大学基础医学院微生物与免疫教研室,郑州市,450052 |
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| 基金项目： | 中国科学院资助项目;河南省教委杰出人才资助项目;河南省郑州市科技攻关项目 |
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| 摘 要： | 目的构建含小鼠MASH1基因的绿色荧光蛋白表达载体pEGFP-C3-MASH1，为进一步研究MASH1在间充质干细胞神经分化中的作用打下基础。方法应用RT-PCR方法从小鼠13．5d胚胎组织中扩增出两端带有HindⅢ和EcoRI酶切位点的MASH1cDNA编码片段，经回收、纯化、酶切后，依次连接到质粒pGEM-T和pEGFP-C3上。结果酶切及测序结果表明：重组质粒PEGFP-C3含有MASH-Ⅰ片段，方向及大小正确。结论成功构建了小鼠真核表达载体PEGFP-C3-MASH-Ⅰ。
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| 关 键 词： | 绿色荧光蛋白 真核表达载体 |
| 文章编号： | 1672-3422（2006）08-0001-04 |
| 收稿时间： | 2006-02-25 |
| 修稿时间： | 2006-02-25 |
Construction of Enhanced Green Fluorescent Protein Eukaryotic Expression Vector Carrying MASH1 Gene |
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| ZHAO Jie ,ZHANG Rui ,XU Yuming ,et al. Construction of Enhanced Green Fluorescent Protein Eukaryotic Expression Vector Carrying MASH1 Gene[J]. Journal of Medical Forum, 2006, 27(8): 1-3,5 |
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| Authors: | ZHAO Jie ZHANG Rui XU Yuming et al |
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| Affiliation: | The First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China |
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| Abstract: | Objective To construct eukaryotic fluorescent expression vector pEGFP-C_3-MASH1 for studing the effect of MASH1 on the neural differentiation of stem cells.Methods The cDNA coding sequence of MASH1 containing the digestion site of Hind III and EcoRI was amplified by RT-PCR from the mouse embryo and then was inserted into pGEM-T plasmid and pEGFP-C_3 fluorescent expression vector.The recombined plasmid was identified by restriction endonuclease digestion?PCR technique and sequencing.Results Restriction erolonuclease digestion and DNA sequencing results showed that the recombinant plasmid PEGFP-C_3 contained the fragment of MASH-I.Conclusion The enkaryotic expression vector PEGFP-C_3-MASH-I was constructed sucessgully. |
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| Keywords: | MASH1 |
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