人粒细胞-巨噬细胞集落刺激因子的克隆及在毕赤酵母中的表达

采用加长引物5＇端的方法克隆了hGM-CSF的编码基因，克隆过程中对该基因的局部做了密码子优化。然后克隆进毕赤酵母分泌型表达载体pPIC9K，电击转化毕赤酵母。PCR、SDS-PAGE与Western blotting证实hGM-CSF已整合进酵母基因组，摇瓶水平粗蛋白表达量达389mg/L，动物实验证实蛋白产物活性正常，SDS-PAGE与N-糖苷酶F去糖基化分析发现hGM-CSF被糖基化。

Cloning and Expression in Pichia pastoris of Human Granulocyte-macrophage Colony Stimulating Factor

Code gene being cloned from the human genome by the primer-5'-extension,as the same time that code had been optimized partly to gene in the cloned procession.It was integrated into the gene secreted Pichia pastoris expression vector pPIC9K and translated yeast Pichia pastoris.PCR,SDS-PAGE and Western blotting confirmed hGM-CSF gene was integrated into the yeast genome and expression was successful.The expressed hGM-CSF was secreted into medium at the level of 389mg/L.Experiments in vivo showed that activity...