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Characterization of regulatory regions involved in the inducible expression of chiB in Bacillus thuringiensis

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Abstract

Expression of the chiB gene from Bacillus thuringiensis Bti75 was defined as inducible by the use of transcriptional fusions with the bgaB reporter gene. The transcription start site of the chiB gene was identified as the C base located 132 base pairs upstream of the start codon. Analysis of 5′ and 3′ deletions of the chiB promoter region revealed that the sequence from position −192 to +36 with respect to the transcription start site was necessary for wild-type levels of inducible expression of the chiB gene. The minimal promoter region for the expression of chiB gene was identified as the sequence from position −100 to +12. Furthermore, a 16-bp sequence (designated dre) downstream of the minimal promoter region of chiB was shown to be required for chitin induction. To confirm the function of this 16-bp sequence, 25 base substitutions were introduced into the dre site. Most of the mutations resulted in constitutive expression, or the efficiency of induction decreased. All mutations identified the dre sequence as a critical site for the inducible expression of chiB. In addition, the dre site was shown to interact with a sequence-specific DNA binding factor of strain Bti75 cultured in the absence of the inducer.

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Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (No. 31371979), the Ph.D. Programs Foundation of Ministry of Education of China (No. 20120031110019) and the Tianjin Natural Science Foundation (No. 12JCYBJC198000).

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  1. Chi-Chu Xie

    Present address: Department of Immunology, Guiyang Medical College, Guiyang, 550004, People’s Republic of China

Authors and Affiliations

  1. Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Tianjin, 300071, People’s Republic of China

    Chi-Chu Xie, Jin Shi, Hai-Yun Jia, Peng-Fei Li, Yang Luo, Jun Cai & Yue-Hua Chen

  2. Department of Microbiology, College of Life Sciences, Nankai University, Weijin Road 94, Tianjin, 300071, People’s Republic of China

    Chi-Chu Xie, Jin Shi, Hai-Yun Jia, Peng-Fei Li, Yang Luo, Jun Cai & Yue-Hua Chen

  3. Tianjin Key Laboratory of Microbial Functional Genomics, Tianjin, 300071, People’s Republic of China

    Jun Cai & Yue-Hua Chen

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Correspondence to Yue-Hua Chen.

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Communicated by Eriko Takano.

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Xie, CC., Shi, J., Jia, HY. et al. Characterization of regulatory regions involved in the inducible expression of chiB in Bacillus thuringiensis . Arch Microbiol 197, 53–63 (2015). https://doi.org/10.1007/s00203-014-1054-3

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  • DOI: https://doi.org/10.1007/s00203-014-1054-3

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