文章摘要
赵 非,黄松明,陆 超,郭 梅,潘晓勤.CD2AP真核表达载体的构建及在COS-7细胞中的表达定位[J].南京医科大学学报,2008,28(1):75~
CD2AP真核表达载体的构建及在COS-7细胞中的表达定位
The construction of eukaryotic expression vector carrying CD2AP and its cellular expressing localization in COS-7 cells
投稿时间:2007-06-11  
DOI:10.7655
中文关键词: CD2AP  绿色荧光蛋白
英文关键词: CD2AP  GFP
基金项目:南京市卫生局科技发展项目(YKK05076)
作者单位
赵 非 南京医科大学附属南京儿童医院肾脏科,江苏 南京 210008 
黄松明 南京医科大学附属南京儿童医院肾脏科,江苏 南京 210008 
陆 超 南京医科大学第一附属医院儿科,江苏 南京 210029 
郭 梅 南京医科大学儿科研究所,江苏 南京 210029 
潘晓勤 南京医科大学儿科研究所,江苏 南京 210029 
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中文摘要:
      目的:构建pEGFP-CD2AP真核表达载体,检测其在COS-7细胞中的表达?方法:PCR扩增CD2相关蛋白(CD2-associated protein,CD2AP)全长编码序列,PCR产物连接入pGEM T-Easy载体,经测序确认无误后,亚克隆入pEGFP-C2构建pEGFP-CD2AP真核表达载体,采用Lipofectamine 2000转染COS-7细胞,荧光显微镜检测报告基因表达产物EGFP?结果:pEGFP-CD2AP表达载体转染COS-7细胞后,可在细胞内观察到报告基因和目的基因的表达产物?结论:pEGFP-CD2AP真核表达载体构建成功,并可在细胞内表达,这将为今后的CD2AP功能研究奠定基础?
英文摘要:
       Objective:To construct the eukaryotic expression vector of pEGFP-CD2AP and detect whether it could express in COS-7 cells. Methods:The coding sequence of CD2AP was obtained by PCR,then the PCR product was insert into pGEM T-Easy vector. After DNA sequence analysis,CD2AP was subcloned into pEGFP-C2 vector. Finally,a eukaryotic expression vector was obtained. The pEGFP-CD2AP expression vector was transfected with lipofectamine 2000 to COS-7 cells,and the expression of pEGFP-CD2AP was determined by GFP fluorescence. Results:The pEGFP-CD2AP expression vector could express both EGFP reporter gene and CD2AP in COS-7 cells. Conclusion:The pEGFP-CD2AP expression vector was constructed successfully,and it could be expressed in COS-7 cells. This work provided foundations for further study on the function of CD2AP.
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