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A truncated derivative of nmt 1 promoter exhibits temperature-dependent induction of gene expression in Schizosaccharomyces pombe

机译:nmt 1启动子的截短衍生物在粟酒裂殖酵母中表现出温度依赖性诱导基因表达

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摘要

Despite increasing exploitation of Schizosaccharomyces pombe as a model system there is a lack of convenient vectors for research and application. Expression with the commonly used promoter, nmt 1, requires a laborious regime involving the removal of repressor, thiamine, from a growing culture and further growth for 18 h to achieve maximum expression, thus underlining the need for more user-friendly promoters. We report here the isolation and characterization of a truncated derivative of the nmt 1 promoter having novel induction characteristics: it is induced by shift of growth temperature from 36 degrees C to 25 degrees C, achieving maximum expression within 3 h. Similar features of expression were observed with the reporter genes GFP and beta-galactosidase, a native gene, cdc 18, and a commercially important foreign therapeutic protein, streptokinase. The new promoter element offers additional advantages, such as lack of deleterious effect on cell viability and potential ability to express toxic proteins. These features make the new promoter a potentially better alternative to nmt 1, both as a research tool and for expression of commercially important proteins in Sz. pombe, and suggest the possibility of using similar approaches to design promoters with novel and useful properties.
机译:尽管越来越多地利用粟酒裂殖酵母作为模型系统,但仍缺乏方便的载体用于研究和应用。使用常用启动子nmt 1进行表达需要费力的方法,包括从生长的培养物中去除阻遏物硫胺素,并进一步生长18 h以实现最大表达,因此强调了对更加用户友好的启动子的需求。我们在这里报告nmt 1启动子的截短的衍生物的分离和表征,具有新的诱导特征:它是由生长温度从36摄氏度变化到25摄氏度诱导的,在3小时内达到最大表达。用报道基因GFP和β-半乳糖苷酶,天然基因cdc 18和商业上重要的外源治疗蛋白链激酶观察到相似的表达特征。新的启动子元件具有其他优点,例如,对细胞生存力无害作用以及表达毒性蛋白的潜在能力。这些功能使新启动子成为nmt 1的潜在更好替代品,无论是作为研究工具还是在Sz中表达商业上重要的蛋白质。 pombe,并建议使用类似方法设计具有新颖和有用特性的启动子的可能性。

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