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

作　　者：徐沁[1] 张丽君[1] 褚孟洋 石菲[2] 闫铭[3] 王永春[2] 

机构地区：[1]第四军医大学学员一旅,陕西西安710032 [2]第四军医大学航空航天医学系航空航天生物动力学教研室,陕西西安710032 [3]第四军医大学西京医院骨科,陕西西安710032

出　　处：《贵州医科大学学报》2017年第6期646-649,共4页Journal of Guizhou Medical University

基　　金：国家自然科学基金项目(81372130;81301681;81301581;81471817)

摘　　要：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：质粒 骨髓间充质干细胞 模拟 失重 过表达载体 satb2 

分 类 号：R34[医药卫生—基础医学]