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Characteristics of the sigK Deletion Mutant from Bacillus thuringiensis var. israelensis Strain Bt-59

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Abstract

All major insecticidal genes of Bacillus thuringiensis var. israelensis (Bti) are controlled by the sporulation-specific sigma factor Sigma E (sigE), while sigE is negatively regulated by Sigma K (sigK). Therefore, knocking out sigK plays an important role in regulating the expression of insecticidal genes in Bti. A sigK deletion mutant of B. thuringiensis var. israelensis strain Bt-59, Bt59(ΔsigK), was constructed by homologous recombination and characterized. The sigK deletion resulted in no mature spores and delayed mother cell lysis from T25 to T60, while the genetically complemented strain, Bt59(HFsigK), had mother cell lysis at T25. Compared to Bt-59, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis indicated that the expression of Cry4Aa2/4Ba1 and Cyt1Aa1 proteins in Bt59(ΔsigK) increased approximately 1.67 and 1.21 times, respectively. However, there was no significant change in Cry11Aa1 protein expression between the two strains. Bioassay results showed that the sigK deletion mutation slightly reduced the insecticidal activity of Bt-59 against Culex pipiens pallens and did not obviously affect activity against Aedes albopictus.

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Acknowledgements

We thank Dr. Didier Lereclus for his critical discussion and suggestions. We also thank Bin Xu, Fan Yang, Na Li, and Jilong Wen for participating in some of the work.

Funding

This work was supported by grants from the National Natural Science Foundation (31530095), the National Key Research and Development Program of China (2017YFD0200400), the Open-end fund of State Key Laboratory for Biology of Plant Diseases and Insect Pests (SKLOF201807), and Jiangsu Province Independent Innovation of Agricultural Science (Grant No. ZX(17)2002).

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  1. Linghuan Xu and Guangjie Han have contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, China

    Linghuan Xu, Xintong Fan, Jing Lv, Xin Zhang, Qi Peng, Jie Zhang & Fuping Song

  2. Department of Applied Microbiology, Jiangsu Lixiahe District Institute of Agricultural Sciences, Yangzhou, 225007, China

    Linghuan Xu, Guangjie Han & Jian Xu

  3. College of Bioscience and Biotechnology, Yangzhou University, Yangzhou, 225002, China

    Linghuan Xu & Jian Xu

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  1. Linghuan Xu
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Xu, L., Han, G., Fan, X. et al. Characteristics of the sigK Deletion Mutant from Bacillus thuringiensis var. israelensis Strain Bt-59. Curr Microbiol 77, 3422–3429 (2020). https://doi.org/10.1007/s00284-020-02150-9

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