Production of an in vitro-derived deletion mutation of Brevibacillus laterosporus by constructing a homology-driven integration vector.
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.
Huang
Wei
W
Engineering Research Center of Industrial Microbiology of Ministry of Education, and College of Life Sciences, Fujian Normal University, Fuzhou, People's Republic of China.
Tian
Bao-Yu
BY
Guo
Jing
J
Cai
Wan-Ling
WL
Huang
Jian-Zhong
JZ
D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D058675:Brevibacillus / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D018274:Electroporation; D005798:Genes, Bacterial; D005822:Genetic Vectors; D010447:Peptide Hydrolases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D012689:Sequence Homology, Nucleic Acid; D012988:Soil Microbiology; D014774:Virulence
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D010447:Peptide Hydrolases