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- W801759813 abstract "设计人Cu，Zn-SOD酵母偏爱密码子并化学合成，与pPIC9K连接，构建酵母偏爱密码子的人Cu，Zn-SOD基因真核表达载体，通过电转化和持续加压筛选毕赤酵母GS115高拷贝转化子，获得的重组高表达酵母菌株建立主种子批。经Southernblot鉴定，基因拷贝数提高2-8倍，活性检测显示提高2-4倍。重组菌的目的基因拷贝数与表达产物呈正相关；表达产物为二聚体，其分子质量为40kD左右，低糖基化，均为分泌表达。Westernblot法分析，对Cu，Zn．SOD抗体具有特异性反应。转化子在培养16h后进入对数生长期，24h后进入生长稳定期；转化子培养20h左右进行诱导表达最为合适。高拷贝的3株重组菌经50次传代后插入的目的基因保持稳定；Cu，Zn．SOD转化子用正交试验筛选摇瓶的诱导表达条件，经诱导表达，Cu，Zn．SOD表达上清最高活性大于600U／mL。确立最适摇瓶培养条件为pH6．0、30℃、体积分数1．5％甲醇诱导72h测得上清的目的蛋白表达最好，表明构建了高表达菌株。" @default.
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- W801759813 date "2012-03-03" @default.
- W801759813 modified "2023-09-24" @default.
- W801759813 title "构建人Cu,Zn-SOD毕赤酵母转化子及其高表达研究" @default.
- W801759813 hasPublicationYear "2012" @default.
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