人凝血因子ⅦcDNA基因的克隆与鉴定

Cloning and Sequencing of Human Coagulatione Factor Ⅶ cDNA Gene from Embryonic Liver

目的:克隆并鉴定人凝血因子ⅦcDNA基因。方法:采用反转录聚合酶链反应(RT-PCR)的方法,从人胎肝总RNA中扩增人凝血因子ⅦcDNA基因,将其克隆入pGEM-T载体,对阳性克隆进行序列测定和分析。结果:经RT-PCR扩增和克隆,获得了人凝血因子ⅦcDNA基因,经序列分析表明,所克隆的基因序列正确。结论:本试验成功克隆了人凝血因子ⅦcDNA基因,为重组人凝血因子Ⅶ的研究奠定了基础。

Coaguladon e factor Ⅶ is a vitamin K dependent glycoprotein which participates in the extrinsic pathway of the blood coagulation.It was concerned for its hemostasis function.In this paper,we extracted the total RNA from human embryonic liver by TRIzol Reagent and amplified the factor Ⅶ cDNA gene using the RT-PCR method, then cloned the gene into pGEM-T vectors.Sequence analysis shows that we have obtained the 1335bp cDNA gene fi＇agment,and the factor Ⅶ cDNA gene was cloned successfully,which is facility to produce the recombination protein for further research.