贵州医科大学学报

2017, v.42;No.201(06) 646-649

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pGV141-satb2真核表达载体的构建及其在骨髓间充质干细胞中的表达
Construction of Satb2 Eukaryotic Expression Vector and Its Expression in Bone Marrow Mesenchyme Stem Cells

徐沁;张丽君;褚孟洋;石菲;闫铭;王永春;
XU Qin;ZHANG Lijun;CHU Mengyang;SHI Fei;YAN Ming;WANG Yongchun;The First Cadet Brigade,Fourth Military Medical University;Department of Aerospace Biodynamics,School of Aerospace Medicine,Fourth Military Medical University;Department of Orthopaedics,Xijing Hospital of Fourth Military Medical University;

摘要(Abstract):

目的:构建satb2的真核过表达载体,观察转入重组载体后C3H10T1/2细胞中satb2的表达情况。方法:根据satb2基因CDS区设计引物,以PCR的方法获得目的基因,将satb2片段双酶切后插入到p GV141载体中进行重组质粒的构建,用PCR和基因测序进行鉴定,将构建成功的重组质粒转染入C3H10T1/2细胞,并通过目的蛋白的检测观察satb2在骨髓间充质干细胞中的表达情况。结果:重组过表达载体GV141-satb2构建成功;与未转染质粒C3H10T1/2细胞相比,转染p GV141-satb2质粒的C3H10T1/2细胞中satb2蛋白表达显著增高,差异有统计学意义(P<0.05)。结论:satb2真核过表达载体构建成功,体外转染实验证实可显著提高骨髓间充质C3H10T1/2细胞中satb2蛋白的表达。
Objective: To construct the eukaryotic expression vector of satb2,and to investigate the expression in bone marrow mesenchyme stem cells( BMSCs) after transfection. Methods: PCR method was employed to obtain the CDS region of satb2. And then the fragment of satb2 was inserted into GV141 vector following by Xho I/EcoRI digestion. The recombinant vector was verified by PCR and DNA sequencing methods. The satb2 expression was detected by western blot after the recombinant vector transfected into BMSCs. Results: p GV141-satb2 was confirmed to be successfully constructed by PCR and DNA sequencing method. Compared with non-transfected group,the expression of satb2 protein was significantly higher in p GV141-satb2 transfection group. Conclusion: The eukaryotic expression vector of satb2 is successfully constructed and can enhance the satb2 expression in BMSCs after transfection.

关键词(KeyWords): 质粒;骨髓间充质干细胞;模拟;失重;过表达载体;satb2
plasmid;bone marrow mesenchyme stem cells;simulation;weightlesshess;overexpression vector;satb2

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基金项目(Foundation): 国家自然科学基金项目(81372130,81301681,81301581,81471817)

作者(Author): 徐沁;张丽君;褚孟洋;石菲;闫铭;王永春;
XU Qin;ZHANG Lijun;CHU Mengyang;SHI Fei;YAN Ming;WANG Yongchun;The First Cadet Brigade,Fourth Military Medical University;Department of Aerospace Biodynamics,School of Aerospace Medicine,Fourth Military Medical University;Department of Orthopaedics,Xijing Hospital of Fourth Military Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2017.06.004

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