用毕赤酵母组成型分泌表达Canstatin-N  

Constitutive and Secreted Expression of Canstatin-N in P. pastoris

作　　者：符策奕 沈锦城[2] 张爱联[2] 罗进贤[3] 张添元[3] 

机构地区：[1]海南省药品检验所,海南海口571101 [2]中国热带农业科学院热带生物技术研究所农业部热带作物学与遗传资源利用重点实验室,海南海口571101 [3]中山大学生命科学学院,广东广州510275

出　　处：《生物技术》2013年第1期19-22,共4页Biotechnology

基　　金：海南省重点科技计划项目("用血管生成抑制因子治疗血管瘤的研究";No.05204);国家自然科学基金项目("血管生成抑制因子治疗鼠癌与人癌差异机理及血管生成抑制因子联合用药研究";No.30760061)资助

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GS115。筛选高G418抗性的克隆作为工程菌GS115(pGAP9K-can-N)。发酵GS115(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56 mg/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。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.

关 键 词：Canstatin—N 毕赤酵母 基因表达 生物活性 

分 类 号：Q786[生物学—分子生物学]