人内皮抑素基因的克隆与序列分析

Cloning and Sequencing of Human Endostatin Gene from Fetal Liver

目的 :克隆与鉴定人内皮抑素基因功能区片段。方法 :采用反转录聚合酶链反应 (RT PCR)的方法 ,从人胎肝总RNA中扩增人内皮抑素基因 ,将其克隆入 pGEM T载体 ,命名为pT hES ,并应用A377自动序列分析仪进行序列分析。结果 :成功克隆了人内皮抑素基因 ,经序列分析表明 ,所克隆的基因序列正确。这为利用基因工程技术生产重组蛋白奠定了基础。

Endostatin, a 20kDa C-terminal fragment of collagen XVIII, is a specific inhibitor for endothelial cell proliferation and angiogenesis. It was concerned for its antitumor activity. In this paper, we extracted the total RNA from human embryonic liver by TRIzol Reagent and amplified the endostatin gene using the RT-PCR method, then cloned the gene into pGEM-T vectors. Sequence analysis shows that we have obtained the 564bp proper gene fragment, and the endostatin gene was cloned successfully, which is facility to produce the recombination protein for further research.