摘要
目的克隆人骨形态发生蛋白-2(hBMP2)基因片段,构建pcDNA3.1-hBMP2真核表达质粒。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态发生蛋白-2基因片段,通过DNA重组技术将该基因片段重组于pcDNA3.1真核表达载体上,构建pcDNA3.1-hBMP2重组质粒,通过用PCR扩增、酶切电泳分析及DNA测序的方法对重组DNA进行鉴定。结果经PCR扩增、酶切电泳分析和DNA测序证实,本实验构建的重组质粒目的基因片段为人BMP2-cDNA。结论本实验成功克隆了hBMP2基因并构建成其真核表达质粒。
Objective To clone human bone morphogenetic protein-2 （hBMP2） gene and construct its eukaryotic expression vector pcDNA3.1-hBMP2. Methods Human BMP2 gene was amplified by RT-PCR method from human osteosarcoma ceils and constructed into eukaryotic expression vector pcDNA3.1-hBMP2. The gene in the vector pcDNA3.1-hBMP2 was identified by PCR amplification, enzyme digestion and DNA sequencing. Results The cloned DNA was confirmed to be hBMP-2 gene. Conclusion In this study, hBMP2 gene is successfully cloned and its eukaryotic expression vector pcDNA3.1-hBMP2 is constructed, which provides the foundation of using BMP2 gene therapy to accelerate new bone formation in distraction osteogenesis.
出处
《华西口腔医学杂志》
CAS
CSCD
北大核心
2007年第5期487-489,共3页
West China Journal of Stomatology
基金
国家自然科学基金资助项目(30160088)
广西壮族自治区自然科学基金资助项目(桂科基0448058)
关键词
骨形态发生蛋白-2
基因克隆
真核表达载体
bone morphogenetic protein-2
gene clone： eukaryotic expression vector
