| 重组pEGFP-C2-p100-TSN点突变质粒构建及表达 |
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| 引用本文： | 高星杰,邵洁,苏超,杨洁. 重组pEGFP-C2-p100-TSN点突变质粒构建及表达[J]. 医学分子生物学杂志, 2010, 7(2). DOI: 10.3870/j.issn.1672-8009.2010.02.010 |
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| 作者姓名： | 高星杰 邵洁 苏超 杨洁 |
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| 作者单位： | 天津医科大学免疫学教研室,天津市,300070;国家教育部免疫微环境与疫病重点实验室,天津市,300070;天津市细胞与分子免疫学重点实验室,天津市,300070 |
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| 基金项目： | 国家高技术研究发展计划(863计划),国家自然科学基金,国家重点基础研究发展规划(973计划),天津市科委国际合作项目,国家自然科学重大研究计划培育项目 |
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| 摘 要： | 目的 将6个不同的p100-TSN.Mutants基因片段分别定向连入PEGFP-C2质粒中,使P100-TSN突变蛋白能够与绿色荧光蛋白在COS7细胞中融合表达,从而为进一步研究p100蛋白TSN结构域的功能奠定实验基础. 方法 利用EcoR Ⅰ和XhoⅠ双酶切方法从6个pcDNA3.1 (+) -p100-TSN.Mutants重组质粒中分别获得p100-TSN.Mutants的cDNA片段,将其连入pEGFP-C2质粒载体中,再将成功构建的6个pEGFP-C2-p100-TSN.Mutants质粒分别转染COS7细胞中,荧光显微镜下观察绿色荧光蛋白表达.结果 ①将重组质粒进行双酶切鉴定可见p100-TSN.Mutants的cDNA片段;②转染重组质粒后可观察到绿色荧光蛋白的表达.结论 ① 6个pEGFP-C2-p100-TSN.Mutants重组质粒构建成功;② p100-TSN突变蛋白可与绿色荧光蛋白在COS7细胞中融合表达.
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| 关 键 词： | 人类P100蛋白 p100-TSN.Mutants pEGFP-C2 重组质粒 融合蛋白 |
Construction and Expression of pEGFP-C2-p100-TSN. Mutants Recombinant Plasmids |
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| GAO Xingjie,SHAO Jie,SU Chao,YANG Jie. Construction and Expression of pEGFP-C2-p100-TSN. Mutants Recombinant Plasmids[J]. Journal of Medical Molecular Biology, 2010, 7(2). DOI: 10.3870/j.issn.1672-8009.2010.02.010 |
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| Authors: | GAO Xingjie SHAO Jie SU Chao YANG Jie |
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| Abstract: | Objective To construct eukaryotic green fluorescence protein expressing recombinant plasmids,pEGFP-C2-p100-TSN.Mutants,which contain human p100-TSN.Mutants cDNAs.Methods The p100-TSN.Mutants cDNAs were purified from EcoRI/XhoI digested pcDNA3.1(+)-p100-TSN.Mutants plasmids and inserted into pEGFP-C2 fluorescent expressing vector. These recombinant pEGFP-C2-p100-TSN.Mutants plasmids were transfected into COS7 cell line,and the expression of green fluorescent fusion proteins was observed by fluorescence microscope.Results ①The fragments of p100-TSN.Mutants were detected by restriction double enzyme digestion. ②The green fluorescent fusion proteins were observed in COS7 cell after the transfection.Conclusion ①The p100-TSN.Mutants fragments were correctly inserted into pEGFP-C2 vector. ②The recombinant pEGFP-p100-TSN.Mutants plasmids effectively expressed GFP-p100-TSN.mutants fusion protein. |
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| Keywords: | p100-TSN.Mutants pEGFP-C2 human p100 protein p100-TSN.mutants pEGFP-C2 recombinant plasmid fusion protein |
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