人内皮抑素基因的克隆与测序

Cloning and sequencing of the fragment of human endostatin gene

目的 获得人内皮抑素 (endostatin)功能区段基因片段 .方法 从人肝组织提取 m RNA,用反转录聚合酶链反应技术克隆 endostatin基因 ,2 0 g· L- 1 脂糖凝胶电泳后将其重组入 p UC19载体 ,双脱氧末端终止法测定其核苷酸序列 .结果 经 Genbank分析证实 ,所获得的 5 5 2 bp基因片段属于endostatin功能区段基因 ,序列正确 .

AIM To procure functional fragment of human endostatin gene. METHODS mRNA from human liver tissue was extracted. And the functional fragment of endostatin gene was amplified by RT PCR. After being identified by 20 g·L -1 agarose gel electrophoresis, it was cloned into pUC19 and sequenced according to dye primer sequencing kit. RESULTS Correct gene fragment of 572 bp was obtained and identified as functional section of endostatin gene by sequencing and Genebank analysis. CONCLUSION Experiment lays foundation for solid tumor antiangiogenesis therapy with endostatin gene.