医学分子生物学杂志2011，Vol.8Issue(2)：105-107,3.DOI:10.3870/j.issn.1672-8009.2011.02.003
小鼠pEGFP-Hoxa11真核表达载体的构建及表达
Construction of pEGFP-Hoxa11 Eukaryotic Expression Vector and Its Expression in CHO Cell Line
摘要
Abstract
Objective To construct the recombinant plasmid pEGFP-Hoxa11 and detect its expression in CHO cell line. Methods The fragments of Hoxa11 was produced by PCR. After enzyme digestion by EcoRI and KpnI, the digested fragments were ligated into pEGFP vector overnight by T4 DNA ligase. The insertion of Hoxa11 in the recombinant plasmid of pEGFP-Hoxa11 was confirmed by PCR, enzyme digestion and DNA sequencing. The recombinant pEGFP-Hoxa11 was transfected into CHO cell lines. and EGFP-Hoxa11 expression was detected by fluorescence microscope and Western blotting analysis. Results The recombinant plasmid pEGFP-Hoxa11 was successfully constructed and its expression was visible in the transfected CHO cells under fluorescence microscope, and Hoxa11 expression was significantly increased in pEGFP-Hoxa11 transfection compared to the endogenous Hoxa11 level in empty vector transfected CHO cells. Conclusion The expression vector pEGFP-Hoxa11 was successfully constructed to co-express Hoxa11 and EGFP protein in CHO cell line.关键词
Hoxa11基因/重组质粒/真核表达分类
医药卫生引用本文复制引用
熊敏,于倩,崔爱娜,张力,朱桂金..小鼠pEGFP-Hoxa11真核表达载体的构建及表达[J].医学分子生物学杂志,2011,8(2):105-107,3.基金项目
国家重点基础研究发展规划(973计划)(No.2007-CB-948104) （973计划）
