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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

作者： Du, Lixin 2 ; Qiu, Lili 1 ; Peng, Qi 1 ; Lereclus, Didier 3 ; Zhang, Jie 1 ; Song, Fuping 1 ; Huang, Dafang 4 ;

作者机构： 1.Chinese Acad Agr Sci, Inst Plant Protect, State Key Lab Biol Plant Dis & Insect Pests, Beijing 100193, Peoples R China

2.IPM Ctr Hebei Prov, Hebei Acad Agr & Forestry Sci, Inst Plant Protect, Baoding, Peoples R China

3.INRA, UMR1319 Micalis, La Miniere, Guyancourt, France

4.Chinese Acad Agr Sci, Biotechnol Res Inst, Beijing 100193, Peoples R China

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.

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