人骨形态发生蛋白-2基因克隆及真核表达载体的构建  

Cloning of human bone morphogenetic protein-2 gene and the construction of its eukaryotic expression vector

作　　者：周诺[1] 黄旋平[1] 廖妮[1] 韦山良[1] 梁飞新[1] 麦华明[1] 

机构地区：[1]广西医科大学附属口腔医院口腔颌面外科,广西南宁530021

出　　处：《华西口腔医学杂志》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)

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.

关 键 词：骨形态发生蛋白-2 基因克隆 真核表达载体 

分 类 号：R78[医药卫生—口腔医学]