突变型hGM-CSF在毕赤甲醇酵母中的分泌表达  

Expression and Secretion of Mutant Recombinant Human GM-CSF in Pichia Pastoris

作　　者：杨红[1] 欧阳克清[1] 郭芝刚[1] 李新平[1] 陈云高 蔡绍皙[1] 

机构地区：[1]重庆大学生物工程学院分子生物学实验室,重庆400044 [2]四川成都荣高实业集团有限公司,成都610000

出　　处：《重庆大学学报（自然科学版）》2002年第6期94-97,共4页Journal of Chongqing University

摘　　要：临床研究发现 ,与大肠杆菌体系生产的hGM -CSF相比 ,利用酵母表达系统生产的hGM -CSF ,具有毒副作用小等优点。鉴于非糖基化的hGM -CSF生物活性比人体天然产生的糖基化GM -CSF活性更高 ,采用PCR定点突变的方法修改hGM -CSF基因中的糖基化位点 ,进而在毕赤酵母 (Pichiapastoris)中实现该突变型基因的高效表达。实验结果表明 ,突变型基因的表达产物具有天然hGM -CSF的活性。Clinic test reveal that by contrasting with expression in E.coli the human granulocyte-macroghage colony-stimulating factor( hGM-CSF) which expression in Pichia Pastoris has lower toxity and side-effect rate. Due to nonglycosylated hGM-CSF has higher biological activity in vivo, we mutant glycosylated positions by using PCR technique. The mutant GM-CSF gene was inserted into PPIC9K and then transformed into Pichia Pastoris strain GS115 for high expression. The results demonstrat that, in spite of the mutation, the expression product also has natural biological activity.

关 键 词：突变型hGM-CSF 毕赤酵母 分泌表达 

分 类 号：Q786[生物学—分子生物学]