中文摘要：

目的：用毕赤酵母的GAP启动子调控组成型表达Canstatin—N。方法：将canstatin—N基因重组于毕赤酵母表达载体pGAP9K的多克隆位点获得pGAP9K—can—N。用电转法将pGAP9K—can—N转化毕赤酵母GSll5。筛选高G418抗性的克隆作为工程菌GSll5（pGAP9K—can—N）。发酵GSll5（pGAP9K—can—N）分泌表达Canstatin—N，用离子交换法纯化目标蛋白。结果：以葡萄糖为碳源，发酵48h分泌表达人血管能抑素蛋白56mg／L。结论：用毕赤酵母的GAP启动子调控组成型表达的人血管能抑素蛋白具有诱发血管内皮细胞凋亡的生物活性。

英文摘要：

Objective： With GAP promoter to control the expression of Canstatin - N in P. pastoris. Method： Canstatin - N gene was cloned on the MCS of pGAP9K to generate of pGAP9K - can - N which was then transformed into the P. pastoris of GSll5 by electroporation method. A strain with higher anti -G418 was used as engineering strain named of GS115 （pGAP9K -can -N）. With glucose as only car- bon source to ferment the strain to express of Canstatin - N and with the method of Ion exchange to purified the aim protein. Result： After 48 h fermentation the GS115 （pGAP9K- can- N） ,56 mg/L of canstatin- N was secreted in the fermentation broth. Conclusion：The re- combinant Canstatin - N expressed heterogeneously in P. pastoris showed activity on induced apoptosis of vascular endothelial cell.