| 小鼠pDsRed-Meis1真核表达载体构建及子宫内表达的鉴定 |
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| 引用本文： | 熊敏,朱桂金.小鼠pDsRed-Meis1真核表达载体构建及子宫内表达的鉴定[J].中国组织化学与细胞化学杂志,2011,20(1):33-35. |
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| 作者姓名： | 熊敏 朱桂金 |
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| 作者单位： | 华中科技大学同济医学院附属同济医院妇产科生殖中心,武汉,430030 |
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| 基金项目： | 国家973基金资助(2007-CB948104) |
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| 摘 要： | 目的构建小鼠Myeloid ectropic viral integration site 1(Meis1)基因和红色荧光蛋白(Red fluorescence protein,RFP)真核表达质粒,为进一步研究Meis1在生殖系统中的功能提供基础。方法设计引物,扩增含Meis1片段的克隆质粒中的meis1基因片段,将其插入真核表达载体pDsRed-N1,重组质粒经酶切鉴定后测序,并转染至小鼠子宫;用Western blot及免疫组织化学方法鉴定外源基因Meis1的表达。结果酶切和测序结果表明,构建的pDsRed-Meis1重组质粒正确;West-ern blot结果显示,外源性Meis1高表达于孕D2的小鼠子宫;转染后冰冻切片结果显示,带红色荧光蛋白的阳性产物表达于孕D4的小鼠子宫内膜及全层;免疫组织化学结果显示,Meis1阳性产物主要表达于孕小鼠子宫上皮、腺体和基质细胞内。结论正确构建了pDsRed-Meis1真核表达质粒,并成功转染及高表达于小鼠子宫,为进一步探讨Meis1在生殖系统中的功能提供了有利工具。
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| 关 键 词： | meis1基因 重组质粒 宫腔内转染 |
Construction of pDsRed-Meis1 expression vector and its expression in murine uterus |
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| Xiong Min,Zhu Guijin.Construction of pDsRed-Meis1 expression vector and its expression in murine uterus[J].Chinese Journal of Histochemistry and Cytochemistry,2011,20(1):33-35. |
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| Authors: | Xiong Min Zhu Guijin |
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| Institution: | Xiong Min,Zhu Guijin(Reproductive Center,Tongij Hospital,Huazhong University of Science and Technology,Wuhan 430030,China) |
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| Abstract: | Objective To construct the recombinant plasmid pDsRed-Meis1 and detect its expression in the murine uterus.Methods The full sequence of Meis1 was obtained by PCR.After enzyme digestion by EcoRI and KpnI,ligation overnight by T4 DNA ligase and transformation by DH5a,we obtained the recombinant plasmid of pDsRed-Meis1,which was identified by enzyme digestion and DNA sequencing.The recombinant plasmid was transfected into the murine uterus,and detected by Western blot.Results The recombinant plasmid pDsRed-Mei... |
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| Keywords: | Meis1 Recombinant plasmid Intrauterus transfection |
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