Ethanol-inducible gene expression using gld1+ promoter in the fission yeast Schizosaccharomyces pombe
Ethanol-inducible gene expression using gld1+ promoter in the fission yeast Schizosaccharomyces pombe
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DOI：
10.1007/s00253-013-4812-2
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发表时间：
2013-08-01
影响因子：
5
通讯作者：
Takegawa, Kaoru
中科院分区：
文献类型：
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作者：
Matsuzawa, Tomohiko;Tohda, Hideki;Takegawa, Kaoru
在裂殖酵母裂殖酵母中，编码甘油脱氢酶的gld 1（+）基因被葡萄糖抑制，并被乙醇和1-丙醇诱导。将gld 1（+）的启动子区克隆到命名为pEG 1的多拷贝载体中，以EGFP作为模型异源蛋白，作为乙醇诱导表达载体进行评估。EGFP的表达在高葡萄糖的存在下被抑制，在乙醇、低葡萄糖和1-丙醇的存在下被诱导，在没有葡萄糖的情况下。向携带pEG 1-EGFP的细胞中加入乙醇被发现是诱导EGFP产生的最有效手段。发现蛋白质产量与乙醇浓度成比例增加。作为有效性的进一步测试，使用pEG 1表达载体在含有甘油和乙醇的培养基中产生分泌的重组人生长激素。pEG 1基因表达系统是在葡萄糖限制条件下生产异源蛋白的有效工具，包括含有甘油作为碳源的培养基。
In the fission yeast Schizosaccharomyces pombe, the gld1 (+) gene encoding glycerol dehydrogenase is repressed by glucose and induced by ethanol and 1-propanol. The promoter region of gld1 (+) was cloned into a multicopy vector designated as pEG1 for evaluation as an ethanol-inducible expression vector using EGFP as a model heterologous protein. Expression of EGFP was repressed in the presence of high glucose and induced in the presence of ethanol, low-glucose, and 1-propanol in the absence of glucose. Addition of ethanol to cells harboring pEG1-EGFP was found to be the most effective means for inducing EGFP production. Protein yields were found to increase in proportion to ethanol concentration. As a further test of effectiveness, secreted recombinant human growth hormone was produced using the pEG1 expression vector in medium containing glycerol and ethanol. The pEG1 gene expression system is an effective tool for the production of heterologous proteins under glucose-limiting conditions, including medium containing glycerol as a carbon source.