人β_2GP1第5功能区cDNA的分子克隆及重组表达

Molecular Cloning and Expression of Human β2 GP1 fifth doman

采用逆转录PCR技术自人胎肝细胞中克隆出人β_2GP1第5功能区cDNA,长度为298bp,经纯化后定向插入表达性载体pGEX-2T,并筛选出带有插入片段的阳性克隆,重组克隆经IPTG诱导,可产生约36kD的融合蛋白,为研究抗磷脂抗体所针对的抗原及表位奠定了基础.

By using reverse transcription-PCR method,the cDNA of the fifth domain of human β_2 GPI with length of 298bp was cloned from neofetal liver cell. The fragment was purified and directionally inserted into the carrier plasmid pGEX-2T. Positive clone containing the inserted fragment were selected and induced by IPTG to produce a fusion protein with 36kD. This study provided solid foundation for further study on an-tiphospholipid antibody-directed antigens and their epitopes.