【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。更多还原

【Abstract】 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.更多还原