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Effect of the spoIIID mutation on mother cell lysis in Bacillus thuringiensis

  • Jing Lv
  • Xin Zhang
  • Tantan Gao
  • Tingting Cui
  • Qi Peng
  • Jie Zhang
  • Fuping SongEmail author
Applied genetics and molecular biotechnology
  • 57 Downloads

Abstract

SpoIIID is a small, sequence-specific DNA-binding protein which can direct many genes’ transcription and has an effect on spore formation in Bacillus subtilis. We investigated the role of SpoIIID in mother cell lysis in Bacillus thuringiensis. A β-galactosidase assay based on the promoter fusions with lacZ indicated that the sigK gene was positively regulated by SpoIIID and σK negatively regulated the expression of sigE. The spoIIID mutant strain exhibited no mother cell lysis in Schaeffer’s sporulation medium (SSM) but did in ½ Luria-Bertani (LB) medium. cwlC is an essential hydrolase gene for mother cell lysis. Moreover, the expression of a PcwlC-lacZ fusion in spoIIID mutant was proved to be higher in ½ LB medium than in SSM. HD (ΔspoIIID)(ΔcwlC) mutant was obtained by knocking out the cwlC gene in HD(ΔspoIIID) and displayed no mother cell lysis in both SSM and ½ LB mediums. The deletion of spoIIID decreased the crystal protein production in HD73. The expression of Porf1cry8E and P5014 promoter fusions with lacZ gene in the acrystalliferous HDspoIIID) mutant showed similar activity to that in the acrystalliferous HD73 strain before T7 and slightly higher than that in the acrystalliferous HD73 after T7. Sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that Cry1Ac production in HDspoIIID) directed by the Porf1cry8E and P5014 promoters was at a similar level as that in HD73 wild strain. Altogether, these results suggested that the spoIIID mutant with Porf1cry8E or P5014 promoters could be an alternative delivery system for cry gene expression with no mature spore formation and medium-dependent mother cell lysis.

Keywords

spoIIID Mother cell lysis cwlC Bacillus thuringiensis 

Notes

Acknowledgments

We thank Dr. Didier Lereclus for his critical discussion and suggestions. We also thank Xiaomin Chen, Linghuan 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 (No. 31530095) and The National Key Research and Development Program of China (No. 2017YFD0200400).

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Jing Lv
    • 1
  • Xin Zhang
    • 1
  • Tantan Gao
    • 1
  • Tingting Cui
    • 1
  • Qi Peng
    • 1
  • Jie Zhang
    • 1
  • Fuping Song
    • 1
    Email author
  1. 1.State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijingChina

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