刘安元,谭立志,万艳平,吴移谋,余敏君,粟盛梅.GFP-hDaxx融合蛋白在真核细胞中的表达与鉴定.[J].中南医学科学杂志.,2003,(1):9-11.
GFP-hDaxx融合蛋白在真核细胞中的表达与鉴定
Expression and Identification of GFP- hDaxx Fusion Protein in Eukaryotic Cells
  
DOI:
中文关键词:  融合蛋白 真核细胞 鉴定 细胞核 GFP-hDaxx hDaxx 早幼粒细胞白血病
英文关键词:hDaxx,fusion protein,eukaryotic cells
基金项目:湖南省教育厅2002年课题资助(NO:02C391).
刘安元  谭立志  万艳平  吴移谋  余敏君  粟盛梅
南华大学病原生物学研究所 湖南衡阳421001 (刘安元,谭立志,万艳平,吴移谋,余敏君)
,南华大学病原生物学研究所 湖南衡阳421001(粟盛梅)
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中文摘要:
      目的:构建GFP-hDaxx融合蛋白真核表达载体pEGFP/hDAXX,并在HeLa细胞中表达,为进一步研究hDaxx在细胞凋亡、病毒感染中的作用提供实验基础。方法:用T4DNA连接酶将hDaxx亚克隆入载体pEGFP,筛选阳性克隆。将质粒pEGFP/hDaxx转染HeLa细胞,Western-Blotting鉴定表达产物,荧光显微镜观察GFP-hDaxx融合蛋白在细胞内的定位。结果:成功构建了GFP-hDaxx融合蛋白真核表达载体pEGFP/hDaxx,GFP-hDaxx融合蛋白成功表达并定位HeLa细胞核。结论:GFP-hDaxx融合蛋白成功地在真核细胞表达并定位于细胞核。
英文摘要:
      Objective To construct eukaryocytic expression vector pEGFP/hDaxx, which can express GFP -hDaxx fusion protein. Methods Subclone hDaxx into pEGFP vector and select positive clone. HeLa cells were transfected with eukaryotic expression vectors encoding GFP and GFP - hDaxx fusion protein. GFP - hDaxx fusion protein was analyzed by Western - blotting and fluoscent localization in HeLa cells. Results pEGFP/hDaxx, which is an eukaryocytic expression vector, was successfully constructed and GFP - hDaxx fusion protein was ideally expressed in HeLa cells, what is more, GFP - hDaxx fusion protein was observed in nuclei under fluoscent microscope . Conclusion GFP - hDaxx fusion protein expressed successfully in eukaryotic cells and localized in nuclei.
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