| 黏附调节因子p120ctn3A细胞核靶向表达质粒的构建及鉴定 |
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| 引用本文： | 李庆昌,戴顺东,王鑫,潘泳岐,徐洪涛,王恩华. 黏附调节因子p120ctn3A细胞核靶向表达质粒的构建及鉴定[J]. 现代肿瘤医学, 2012, 20(2): 234-238. DOI: 10.3969/j.issn.1672-4992.2012.02.06 |
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| 作者姓名： | 李庆昌 戴顺东 王鑫 潘泳岐 徐洪涛 王恩华 |
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| 作者单位： | 中国医科大学基础医学院病理教研室,附属第一医院病理科,辽宁,沈阳,110001 |
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| 基金项目： | 国家自然科学基金资助项目(编号:30700806,81071717) |
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| 摘 要： | 目的 既往少数研究表明p120ctn存在核浆穿梭现象,为研究细胞核内p120ctn的功能,我们试图构建p120ctn 3A细胞核定向表达载体pCMV/p120ctn 3A.方法 采用PCR方法扩增人p120ctn 3A活性片段,将其插入细胞核定向表达载体pCMV/myc/nuc,构建重组体pCMV/p120ctn 3A,经脂质体介导转染肺癌细胞A549,用荧光显微镜观察绿色荧光信号,免疫细胞化学及Western blot鉴定其在肺癌细胞A549中的表达与定位.结果 限制性双酶切及DNA测序分析结果显示插入pCMV/p120ctn 3A的片段为2496bp左右,与预期p120ctn 3A基因片段大小相同.荧光显微镜观察到绿色荧光信号定位于细胞核内,免疫细胞化学结果显示表达的p120ctn 3A定位于A549细胞核.Western blot结果显示转染pCMV/p120ctn 3A的A549细胞p120ctn 3A蛋白表达量显著上调.结论 成功构建了p120ctn 3A 细胞核定向表达载体pCMV/p120ctn 3A.
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| 关 键 词： | p120ctn 核内定位载体pCMV/myc/nuc 重组质粒 |
Construction and identification of p120ctn 3A nucleus-localization plasmid |
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| LI Qingchang,DAI Shundong,WANG Xin,PAN Yongqi,XU Hongtao,WANG Enhua. Construction and identification of p120ctn 3A nucleus-localization plasmid[J]. Journal of Modern Oncology, 2012, 20(2): 234-238. DOI: 10.3969/j.issn.1672-4992.2012.02.06 |
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| Authors: | LI Qingchang DAI Shundong WANG Xin PAN Yongqi XU Hongtao WANG Enhua |
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| Affiliation: | Department of Pathology,College of Basic Medical Sciences and 1st Affiliated Hospital,China Medical University,Liaoning Shenyang 110001,China |
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| Abstract: | Objective:To construct p120ctn 3A nuclear target localizaiton plasmid.Methods: p120ctn 3A cDNA was amplified by PCR and inserted to nucleus-localization expression vector pCMV/myc/nuc to construct pCMV/p120ctn 3A.Then the pCMV/p120ctn 3A was transfected into A549 cells.The expression and localization of p120ctn 3A in A549 cells were verified by fluorescent microscope,immunocytochemistry and Western blot.Results: The size of inserted fragments in the recombinant vectors pCMV/p120ctn 3A was 2496bp,corresponding to that of p120ctn 3A.After transfecting pCMV/M-CSF into A549 cells by Lipofectamine 2000,pCMV/p120ctn 3A-transfected A549 cells could express p120ctn 3A protein in nucleus.The expression of p120ctn 3A was significantly elevated in pCMV/p120ctn 3A-transfected A549 cells.Conclusion: The nuclear localization vector pCMV/p120ctn 3A was successfully constructed. |
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| Keywords: | p120ctn nuclear localization vector pCMV/myc/nuc recombinant plasmid |
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