人canstatin基因的克隆

Cloning of human canstatin gene

为深化研究血管生成抑制剂canstatin的生物学性能和用于肿瘤生物治疗的可能性,针对canstatincDNA序列设计引物,运用逆转录聚合酶链反应(RT-PCR)从人胚肝组织中钓取canstatincDNA,将其克隆入pMD18-T载体中,通过酶切鉴定出重组体并测序分析。结果表明获得684bp人canstatin基因,成功构建人canstatincDNA克隆载体pMD18-T/canstatin,为进一步进行canstatin蛋白表达及活性研究奠定了基础。

For researching biological activities and potential utility of canstatin as an inhibitor of angiogenesis,canstatin gene cDNA fragment was amplified from total RNA extracted from fresh fetal liver by RT-PCR.The re-combinant plasmid pMD18-T/canstatin was constructed by inserting the canstatin cDNA into cloning vector pMD18-T and transformed into E.coli DH5α.Indentify the positive cloning that contain the cDNA of canstatin by endonu-clease digestion and sequencing confirmed the obtain cDNA fragment was consistent with published sequence.Thus laying the foundation of further study on expression and protein biological activities.