人腺激肽释放酶2在毕赤酵母中的表达及活性测定  

Expression of Human Glandular Kallikrein 2 by Pichia Pastoris (Yeast) and its Activity Assay

作　　者：路英丽[1] 赵慧[2] 邹冬辉[3] 李丹地[3] 郭焱[3] 王忠山[3] 

机构地区：[1]吉林大学第二医院,长春130041 [2]吉林大学基础医学院组织胚胎学教研室,长春130021 [3]吉林大学基础医学院细胞生物学教研室,长春130021

出　　处：《生殖与避孕》2007年第7期433-437,共5页Reproduction and Contraception

基　　金：国家自然科学基金资助项目(No.39870313)

摘　　要：目的:利用在巴氏毕赤酵母细胞表达重组人腺激肽释放酶2(rhK2)。方法:构建表达载体pPICZαC-mhK2及具有3个表达盒子的pPICZαC-3mhK2。采用电转法,将重组表达质粒转染毕赤酵母X33细胞,甲醇诱导表达,离子交换层析纯化rhK2蛋白;检测rhK2的酶活性。结果:获得了高表达rhK2的酵母细胞株,其酶活性接近正常精浆中hK2的活性。结论:毕赤酵母能够成功表达具有天然活性的人腺激肽释放酶2(hK2)。Objective： To investigate the expression of recombinant human glandular kallikrein 2 in the methylotropic yeast Pichia pastorsi. Methods： The expression vectors pPICZαC-mhK2 and multimers pPICZαC- 3mhK2 which have 3 expression cassettes were constructed and transferred into X33 cells by electroporation. The protein was expressed in Pichia pastorsi induced with methanol. The rhK2 was purified by anion-exchange chromatography and its activity was assayed. Result： rhK2 was obtained in Pichia pastorsi with its biological activity is almost equal to the natural hK2. Conclusion： The mature rhK2 can be expressed in Pichia pastoris.

关 键 词：人腺激肽释放酶2(hK2) 毕赤酵母 

分 类 号：Q78[生物学—分子生物学]