报告基因法比较两种放线菌启动子的活性

Evaluation of the activities of two promoters in Streptomycetes by reporter gene method

【目的】比较启动子Psf与红霉素抗性基因启动子(PermE*)在链霉菌中的表达强度差异。【方法】本文利用卡那霉素抗性梯度以及邻苯二酚2,3-双加氧酶显色系统,比较了两个启动子的表达差异。【结果】两个启动子在棒状链霉菌(Streptomyces clavuligerus)NRRL3585、天蓝色链霉菌(Streptomyces coelicolor)M145,委内瑞拉链霉菌(Streptomyces venezuelae)ISP5230及变铅青链霉菌(Streptomyces lividansTK54)均高表达。在棒状链霉菌和天蓝色链霉菌中Psf的表达活性要强于PermE*。【结论】Psf和PermE*是适用于链霉菌的高活性启动子。这些数据为两个启动子的使用提供了参考依据,也为链霉菌基因表达元件提供了新的选择。

[ Objective] The mutated promoter of the erythromycin resistance gene（ PermE ＊ ） is a strong promoter generally used in streptomycetes, and we evaluated the expression activities of a new promoter（Psf） and PermE＊ in Streptomycetes. [Methods] We used kanamycin resistance gene（neo） and catechol 2,3-dioxygenase gene（xylE） as reporters. [Results] Both promoters exhibited high level of promoter activities in Streptomyces clavuligerus NRRL3585, Streptomyces coelicolor M145, Streptomyces venezuelae ISP5230 and Streptomyces lividans TK54. The activities of Psf were higher than those of PermE ＊ in S. clavuligerus and S. coelicolor. [ Conclusion] Both Psf and PermE ＊ are strong promoters suitable for gene over-expression in Streptomycetes and Psf will offer an alternative for high-level gene expression.