Optimization of Electroporation Conditions for Bacillus pumilus 3–19 Strain

Danilova, Iuliia V.; Рудакова, Н. Л.; Vasilyeva, Y. A.; Gilmutdinova, A. I.; Diadkina, I. V.; Khasanov, D. I.; Шарипова, М. Р.

doi:10.1007/s12668-022-00982-2

Cited by 1 publication

(2 citation statements)

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“…The resulting plasmids (pDIb11.21, pVYb11.21, and pGAs11.21) were transformed into B. pumilus 3-19 cells using an optimized electroporation method [ 22 ]. Transformant colonies are shown in Figure S3 .…”

Section: Resultsmentioning

confidence: 99%

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Characterization of Bacillus pumilus Strains with Targeted Gene Editing for Antimicrobial Peptides and Sporulation Factor

Danilova,

Vasileva,

Gilmutdinova

et al. 2023

Microorganisms

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Due to their capacity to produce antimicrobial peptides that can prevent the growth of diseases, many Bacillus spp. are beneficial to plants. In this study, we looked into the antagonistic activity of the B. pumilus 3-19 strain and its derivatives following targeted genome editing. Two peptide genes with antibacterial action, bacilysin (bac) and bacteriocin (bact), and the sigF gene, which encodes the sigma factor of sporulation, were specifically inactivated using the CRISPR-Cas9 system in the genome of B. pumilus 3-19. Antibacterial activity against B. cereus and Pantoea brenneri decreased as a result of the inactivation of target genes in the B. pumilus 3-19 genome, with a noticeable effect against bacilysin. The growth dynamics of the culture changed when the bac, bact, and sigF genes were inactivated, and the altered strains had less proteolytic activity. An asporogenic mutant of B. pumilus 3-19 was obtained by inactivating the sigF gene. It has been proven that bacilysin plays a unique part in the development of B. pumilus 3-19’s antagonistic action against soil microorganisms.

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Section: Resultsmentioning

confidence: 99%

“…Transformation of recombinant DNA into E. coli DH5α cells was performed using CaCl2 [ 21 ]. B. pumilus 3-19 cells were transformed using AEB1 electroporation buffer pH 6.0 [ 22 ].…”

Section: Methodsmentioning

confidence: 99%