Identification and transcriptional analysis of a cold shock-inducible gene, cspA, in Streptomyces coelicolor A3(2).

J Kormanec, B Sevcíková
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引用次数: 19

Abstract

Using the method for identification of promoters recognized by the sporulation-specific sigma factor sigmaF, we identified a promoter, cspAp, in Streptomyces coelicolor, which showed similarity to the consensus sequence of Bacillus subtilis promoters recognized by the general stress-response sigma factor sigmaB. cspAp directs expression of the cspA gene, which shows sequence similarity to members of the family of major cold shock proteins (CspA) from bacterial species. S1-nuclease mapping using RNA prepared from Escherichia coli containing a two-plasmid system, and from Streptomyces coelicolor at various developmental stages, identified identical transcription start points in both species, corresponding to cspAp. However, the promoter was also active in a S. coelicolor sigF mutant. Transcriptional studies indicated that cspA is transcribed as a monocistronic mRNA. The level of cspA mRNA remains almost constant in all developmental stages, is dramatically increased after cold shock, and decreased after heat shock. Disruption of the S. coelicolor cspA gene did not affect growth or differentiation at 11 degrees C or 30 degrees C.

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冷冲击诱导基因cspA的鉴定和转录分析(2)。
利用孢子特异性sigma因子sigmaF识别启动子的方法,我们在链霉菌中鉴定出一个启动子cspAp,该启动子序列与枯草芽孢杆菌中通用应激反应sigma因子sigmaB识别的启动子序列相似。cspAp指导cspA基因的表达,其序列与细菌主要冷休克蛋白(cspA)家族成员相似。利用含有双质粒系统的大肠杆菌和处于不同发育阶段的彩色链霉菌制备的RNA对s1 -核酸酶进行定位,鉴定出两种物种中相同的转录起点,对应于cspAp。然而,该启动子在S. colcolcolor sigF突变体中也有活性。转录研究表明,cspA转录为单顺反子mRNA。cspA mRNA水平在各发育阶段基本保持不变,冷休克后显著升高,热休克后显著降低。在11℃和30℃条件下,对S. coelicolor cspA基因的破坏不影响其生长和分化。
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