摘要
目的:构建pcDNA3.1-hBMP2真核表达质粒并检测其在兔骨髓间充质干细胞(BMSCs)中的表达。方法:采用逆转录聚合酶链式反应(RT-PCR)技术,从人骨肉瘤中扩增出人骨形成蛋白-2(hBMP2)基因片段,构建成pcDNA3.1-hBMP2重组质粒。转染兔骨髓间充质干细胞并通过RT-PCR和免疫组化检测其表达。结果:本实验构建的重组质粒目的基因片段为hBMP2-cDNA。经RT-PCR和免疫组化证实,转染pcDNA3.1-hBMP2后的兔骨髓间充质干细胞内有大量hBMP2 mRNA的转录和蛋白的表达。结论:本实验成功构建hBMP2真核表达质粒并在骨髓间充质干细胞中得到表达,为进一步研究用BMP2基因转染的方法来加速牵张成骨新骨形成奠定了基础。
Objective： To construct human bone morphogenetic protein-2 （hBMP2） eukaryotic expression vector and detect its expression in rabbit＇s bone mesenchymal stem cells. Methods： Human BMP2 gene was amplified by RT-PCR method from human osteoma cells and constructed into eukaryotic expression vector pcDNA3.1- hBMP2. Its expression was detected by RT-PCR and immunohistochemical analysis after transfection rabbit＇s bone mesenchymal stem cells. Result： The cloned DNA was confirmed to be hBMP2 gene. Its expression was confirmed by RT-PCR and immunohistochemical analysis after transfection of rabbit＇s bone mesenchymal stem cells. Conclusion： In this study we successfully constructed BMP2 eukaryotic expression vector and BMP2 could be expressed in bone mesenchymal stem cells after transfection, which provided the foundation of using BMP2 gene therapy to accelerate new bone formation in distraction osteogenesis.
出处
《广西医科大学学报》
CAS
北大核心
2007年第3期338-341,共4页
Journal of Guangxi Medical University
基金
国家自然科学基金资助项目(No30160088)
广西自然科学基金资助项目(No桂科基0448058)
广西区教育厅项目(No200510023)
广西卫生厅项目(No桂卫重200525)
