不同猪胰岛素前体基因拷贝数Mut^+型毕赤酵母的摇瓶发酵研究  

Investigation on multiple porcine insulin precursor gene copy Pichia pastoris Mut^+ strains by shake flask

作　　者：张明[1] 郭美锦[1] 庄英萍[1] 储炬[1] 张嗣良[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室国家生化工程技术研究中心,上海200237

出　　处：《工业微生物》2010年第2期35-39,共5页Industrial Microbiology

基　　金：国家自然科学基金资助项目(30560006)

摘　　要：毕赤酵母是优秀的外源蛋白的表达系统之一。本文对Mut^+型的不同PIP基因拷贝数的毕赤酵母进行了摇瓶试验。研究了生长特性以及对外源蛋白表达量的影响等。发现了低拷贝和高拷贝的蛋白表达量、生长情况有差异。G12重组菌的PIP表达量最高为181.6mg/L,是单拷贝重组菌表达量的12.6倍。对于基因拷贝数低于12的菌株,PIP表达水平与PIP基因拷贝数成线性关系(r=0.996)。The methylotrophic yeast, Pichia Pastoris, was one of most widely used expression system for heterologous proteins production in the past decades. The shaking flask cultivation was used to research the growth specificity and PIP production of multi-copy Pichia pastoris strains. Difference between high and low gene dosage strain was found. The PIP production of G12 was 181.6mg/L, 12 times than that of the single-copy stain. It was found that PIP production positively related with gene dosage when the copy numbers were less than 12 and the correlation coefficient was 0. 996.

关 键 词：重组毕赤酵母 基因拷贝数 猪胰岛素前体 

分 类 号：Q78[生物学—分子生物学]