Functional identification of the gene bace16 from nematophagous bacterium Bacillus nematocida.
Bacillus nematocida is a Gram-positive bacterium capable of killing nematodes. Our recent studies identified an extracellular serine protease Bace16 in B. nematocida as a candidate of pathogenic factor in the infection against nematodes, which displayed a high similarity with the serine protease family subtilisin BPN', and the MEROPS ID is S08.034. To further confirm the roles that bace16 played in the mechanism of nematocidal pathogenesis, recombinant mature Bace16 (rm-Bace16) was expressed in Escherichia coli strain BL21 using pET-30 vector system. Bioassay experiments demonstrated that the purified recombinant protease had the ability to degrade nematode cuticles and kill nematodes. In addition, a bace16 knockout mutant of B. nematocida constructed by homologous recombination showed considerably lower proteolytic activity and less than 50% nematocidal activity than the wild-type strain. These results confirmed that Bace16 could serve as an important virulence factor during the infectious process.
Niu
Qiuhong
Q
Laboratory for Conservation and Utilization of Bio-Resources, Yunnan University, Kunming, PR China.
Huang
Xiaowei
X
Zhang
Lin
L
Lian
Lihui
L
Li
Yunxia
Y
Li
Juan
J
Yang
Jinkui
J
Zhang
Keqin
K
D000818:Animals; D001407:Bacillus / Q000235:genetics / Q000472:pathogenicity*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001681:Biological Assay; D017353:Gene Deletion; D009348:Nematoda / Q000382:microbiology*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism*; D014774:Virulence
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D012697:Serine Endopeptidases