人骨保护素编码区基因的克隆及其真核表达载体的构建  

Cloning of human osteoprotegerin and construction of mammalian expression vector pcDNA3.1-OPG-IRES-EGFP

作　　者：王晋东[1] 王岩[1] 周勇刚[1] 王冉东[1] 俞广[1] 白金柱[1] 黄鹏[1] 张国强[1] 柴伟[1] 

机构地区：[1]中国人民解放军总医院骨科医院,北京100853

出　　处：《山西医科大学学报》2007年第8期673-676,共4页Journal of Shanxi Medical University

摘　　要：目的克隆人骨保护素(OPG)编码区基因并构建其真核表达载体。方法以人骨肉瘤细胞系MG63总RNA为模板,采用RT-PCR方法获取OPG编码区基因核苷酸序列,应用定向克隆技术,构建真核表达载体pcDNA3.1-OPG-IRES-EGFP。结果获取的OPG编码区全长核苷酸序列与文献报道的完全一致,经PCR和多酶切鉴定证实构建的表达载体正确。结论获得了人骨保护素(OPG)编码区基因,并成功构建了其真核表达载体。Objective To clone the encoding gene of human osteoprotegerin（hOPG）and construct the eukaryotic expression vector pcDNA3.1-OPG-IRES-EGFP.Methods Using the isolated total RNA from osteosacoma cell line MG63 as a template,the cDNA encoding hOPG was amplified by reverse transcription-polymerase chain reaction method.Further,the PCR product was cloned into pGEM-T Easy and sequenced.Finally,Recombinant vector pcDNA3.1-OPG-IRES-EGFP was constructed by directionally cloning the target gene fragments into eukaryotic expression vector pcDNA3.1（＋）.Results The OPG cDNA was identical with related literatures,and the recombinant vector pcDNA3.1-OPG-IRES-EGFP was verified by extensive restriction digests and PCR.Conclusion The OPG cDNA is obtained and the recombinant plasmid is constructed successfully.

关 键 词：骨保护素 基因克隆 载体构建 

分 类 号：Q782[生物学—分子生物学]