Abstract
In this study, a homology-driven integration vector and electroporation system was developed to delete a protease gene in the pathogenic bacterium Brevibacillus laterosporus strain G4. Furthermore, an in vitro protease-deficient mutation was generated by introducing the integration vector with a 445-bp protease BLG4 fragment into B. laterosporus chromosomal target via homologous recombination. The BLG4-deficient mutant showed a significant drop in protease activity as compared to the wild-type G4 strain, but had a slight effect on bacterial growth and sporulation. The results revealed that the developed method can become an important tool for studying the molecular pathogenesis mechanisms of B. laterosporus.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Bacterial Proteins / genetics
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Bacterial Proteins / isolation & purification
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Bacterial Proteins / metabolism
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Base Sequence
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Blotting, Western
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Brevibacillus / enzymology
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Brevibacillus / genetics*
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Brevibacillus / metabolism
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Brevibacillus / pathogenicity
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Cloning, Molecular
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DNA, Bacterial / genetics
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Electroporation
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Genes, Bacterial
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Genetic Vectors*
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Peptide Hydrolases / genetics*
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Peptide Hydrolases / isolation & purification
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Peptide Hydrolases / metabolism
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Plasmids / genetics
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Recombination, Genetic
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Sequence Analysis, DNA
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Sequence Deletion*
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Sequence Homology, Nucleic Acid*
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Soil Microbiology
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Virulence
Substances
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Bacterial Proteins
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DNA, Bacterial
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Peptide Hydrolases