神经损伤与功能重建2011，Vol.6Issue(4)：235-240,6.DOI:10.3870/sjsscj.2011.04.001
携带人类GAD67基因真核表达载体的构建及转染骨髓基质细胞
To Construct Eukaryotic Expression Vector with GAD67 Gene and Transfect It into Bone Marrow Stromal Cells
摘要
Abstract
Objective: To construct human glutamic acid decarboxylase (GAD67) gene's recombinant eukaryotic expression vector pDC316-CMV-EGFP-GAD67 and to transfect it into bone marrow stromal cells (BMSCs) of rats. Methods: Eukaryotic expression vector pDC316-CMV-EGFP-GAD67 was constructed with the technique of gene rearrangement. BMSCs of rats were cultivated by their adherence-dependent growth character, and the recombinant pDC316-CMV-EGFP-GAD67 was transferred into the BSMCs with lipofectamineTM2000. The cells were observed by fluorescence microscopy, the efficiency of transfection was assessed by flow cytometry, and the expression of GAD67 protein in BMSCs was detected by immunohistochemistry after transfecting. Results: The pDC316-CMV-EGFP-GAD67 was constructed successfully. Green fluorocytes were detected under fluorescence microscopy after transfection and the efficiency of transfection was 37.30％ at 48 hours post transfection. GAD67 protein expressed in tansfected BMSCs at 48 hours post transfection. Conclusion: The eukaryotic expression vector pDC316-CMV-EGFP-GAD67 was successfully constructed and effectively transfected into BMSCs.关键词
谷氨酸脱羧酶67/真核表达载体/转染/骨髓基质细胞Key words
glutamic acid decarboxylase67/ eukaryotic expression vector/ transfection/bone marrow stromal cells分类
生物科学引用本文复制引用
谢三平,陈吉相,李慧,徐婧,黄园园..携带人类GAD67基因真核表达载体的构建及转染骨髓基质细胞[J].神经损伤与功能重建,2011,6(4):235-240,6.基金项目
湖北省科技攻关计划项目(No.GGS0064) （No.GGS0064）
