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重组Pegfp-C2-PSF质粒的构建及表达 被引量：1

Construction and Expression of pEGFP-C2-PSF Recombinant Plasmid
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摘要 目的将人类PSF基因定向连人pEGFP-C2质粒，使PSF蛋白可与绿色荧光蛋白在HeLa细胞内融合表达，为进一步研究PSF蛋白的功能及定位奠定实验基础。方法采用EcoR I和Sal I双酶切方法，从pGEX-2TK-PSF质粒中获得PSF蛋白的cDNA全长；连入pEGFP-C2质粒的C端。将构建成功的pEGFP-C2-PSF质粒转染人HeLa细胞，荧光显微镜下观察绿色荧光蛋白表达。结果①将该质粒进行双酶切鉴定可见PSF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①PSFcDNA全长成功连入pEG-FP-C2质粒；②PSF蛋白可与绿色荧光蛋白在HeLa细胞中融合表达。 Objective To construct eukaryotic green fluorescent protein （GFP） expressing recombinant plasmids, pEGFP-C2-PSF, which contains human PSF full length cDNA. Methods The PSF full length cDNA was obtained from EcoR I/Sal I digested pGEX-2TK-PSF plasmid, and inserted into pEGFP-C2 fluorescent expressing vector. This recombinant pEGFP-C2-PSF plasmid was transfected into HeLa cell line and the expression of green fluorescent fusion protein was observed by fluorescence microscopy. Results （1） The PSE fragment was seen after double digestion of pEGFP-C2-PSF. （2） The expression of green fluorescent fusion protein was detected in the HeLa cells transfected with recombinant pEGFP-C2-PSF. Conclusion The full length of PSF cD- NA fragment was correctly inserted into the pEGFP-C2 vector. The fluorescent expressing recombinant plasmids pEGFP-C2-PSF expressed PSF-GFP fusion proteins successfully.
作者 东莉洁 李晓冬 何津岩 姚智 杨洁
机构地区 天津医科大学免疫学教研室 天津市细胞与分子免疫学重点实验室
出处 《医学分子生物学杂志》 CAS CSCD 2009年第3期235-238,共4页 Journal of Medical Molecular Biology
基金 国家高技术研究发展计划（863计划）（No.2007AA02Z115）,国家自然科学基金（No.30670441,30300070）,国家教育部新世纪人才支持计划（No.NCET-04-0245）,天津市科委应用基础研究重点项目（No.07JCZDJC07300）,天津市科委国际合作项目（No.05YFGHHZ01300）
关键词 人类PSF蛋白 PEGFP-C2 重组质粒 融合蛋白 Human PSF protein pEGFP-C2 Recombinant plasmid Fusion protein
分类号 Q522.2 [生物学—生物化学]
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参考文献10

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同被引文献20

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医学分子生物学杂志
2009年 第3期

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