小鼠内皮抑素基因的克隆及序列测定

Cloning and sequencing of mouse endostatin gene

目的 获取小鼠内皮抑素 (endostatin)基因并进行序列测定 ,为今后研究其机制及应用创造条件 .方法 以小鼠肝脏总 RNA为模板 ,用 RT- PCR法从中扩增 endostatin基因 ,并将所得基因重组入 p UC19载体质粒 ,采用自动测序仪及荧光素标记引物 ,测定目的基因序列 .结果 从小鼠肝组织中成功扩增 endostatin基因 ,并成功克隆入 p U C19载体 .经酶切鉴定正确后 ,命名为 p UC- Endo.经测序证实所获基因序列与已知的 endostatin基因序列一致 .结论 成功构建小鼠 endo-statin基因的重组克隆 p U C- Endo,为今后利用

AIM To obtain mouse endostatin gene and to explore its mechanism and application. METHODS cDNA encoding mouse endostatin was amplified from mouse liver by RT PCR and cloned into the plasmid pUC19. The nucleotide sequence of the target gene was determined by the Perkin Elmer Amplied Biosystems 373 Sequencer with positive and reverse primers. RESULTS Endostatin gene clone was successfully constructed and named pUC-Endo. And the gene sequence cloned into pUC19 was consistent with the known sequence after determination. CONCLUSION The recombinant mouse endostatin gene clone has been establishedsuccessfully and has provided a basis for further research of the glioma anti vessel therapy.