Effect of the spoIIID mutation on mother cell lysis in Bacillus thuringiensis
Autor: | Tingting Cui, Jing Lv, Fuping Song, Xin Zhang, Jie Zhang, Peng Qi, Tantan Gao |
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Rok vydání: | 2018 |
Předmět: |
Lysis
Mutant Bacillus thuringiensis lac operon Bacillus subtilis Applied Microbiology and Biotechnology 03 medical and health sciences Hemolysin Proteins Bacteriolysis Bacterial Proteins Transcription (biology) Gene expression Gene 030304 developmental biology Spores Bacterial 0303 health sciences biology Bacillus thuringiensis Toxins 030306 microbiology Chemistry fungi Promoter General Medicine Gene Expression Regulation Bacterial biology.organism_classification Molecular biology DNA-Binding Proteins Endotoxins Gene Deletion Biotechnology Transcription Factors |
Zdroj: | Applied microbiology and biotechnology. 103(10) |
ISSN: | 1432-0614 |
Popis: | 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 HD-(ΔspoIIID) 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 HD-(ΔspoIIID) 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. |
Databáze: | OpenAIRE |
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