Identification of the promoter in the intergenic region between orf1 and cry8Ea1 controlled by sigma H factor

文献类型: 外文期刊

第一作者: Qiu, Lili

作者: Qiu, Lili;Peng, Qi;Zhang, Jie;Song, Fuping;Du, Lixin;Lereclus, Didier;Huang, Dafang

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期刊名称:APPLIED AND ENVIRONMENTAL MICROBIOLOGY ( 影响因子:4.792; 五年影响因子:5.26 )

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收录情况: SCI

摘要: Bacillus thuringiensis Cry8Ea toxin is specifically toxic to larvae of the Asian cockchafer, Holotrichia parallela. Here we investigated the mechanism of transcriptional regulation of the cry8Ea1 gene. Reverse transcription-PCR (RT-PCR) results indicated that cry8Ea1 and an upstream gene (orf1) were cotranscribed. Transcriptional fusions with the lacZ gene demonstrated that transcription of the cry8Ea1 gene started from two promoters: P _(orf1), which is located upstream of the orf1 gene, and P _(cry8E), located in the intergenic region mapping between orf1 and cry8Ea1. Of the known, similar orf1-cry operons, this is the first report of the existence of a promoter in the intergenic region between the orf1 and cry genes. The transcriptional activity of P _(orf1) was found during sporulation in B. thuringiensis subsp. kurstaki HD-73 and was almost abolished in the sigE mutant, while the transcriptional activity of P _(cry8E) was detected after the end of the exponential phase in HD-73 and was considerably lower in the sigH mutant. The transcription start sites generated by the two cry8Ea1 promoters were determined by the 5' -SMARTer rapid amplification of cDNA ends (RACE) method. The-35 and-10 regions of P _(orf1) and P _(cry8E) showed high sequence similarity with the σ ~E and σ ~H promoters, respectively. These results indicated that P _(orf1) is controlled by the σ ~E factor and P _(cry8E) by the σ ~H factor.

分类号: Q939.9

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