小鼠血管抑素基因的克隆及真核表达载体构建

Cloning of Mouse Angiostatin and Construction of its Eukaryotic Expression Vector

目的 :克隆小鼠血管抑素 (Angiostatin)cDNA ,构建其真核表达载体pCMV -Angiostatin重组质粒 ,为进一步研究其抗肿瘤作用奠定基础。方法 :根据Genebank中小鼠血管抑素基因序列 ,用RT -PCR方法从鼠肝脏中扩增出AngiostatincDNA ,连接 pMD18T载体测序 ,经测序证实后 ,通过中间载体 pKS ,构建pCMV -Angiostatin重组质粒。结果 :测序表明扩增的AngiostatincDNA序列与报道基本一致 ,AngiostatincDNA正确插入表达载体。 结论

Objective: To clone the sequence of mouse Angiostatin cDNA and construct its eukaryotic expression vector, provide a base for its antitumor effect. Methods: According to the sequence of Mouse angiostatin cDNA in Genebank, Mouse angiostatin cDNA was amplified by RT-PCR and ligated into pMD18T vector. Through the vector pKS, the pCMV-Angiostatin recombinant plasmid was constructed. Results: The sequence of mouse Angiostatin cDNA is identical with reported. Angiostatin cDNA was inserted into expression vector correctly. Conclusion: Mouse Angiostatin cDNA is cloned and pCMV-Angiostatin recombinant plasmid is completed successfully.