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Genet Mol Res ; 14(3): 10619-29, 2015 Sep 09.
Article in English | MEDLINE | ID: mdl-26400293

ABSTRACT

Biofilm-forming bacteria are highly resistant to antibiotics, host immune defenses, and other external conditions. The formation of biofilms plays a key role in colonization and infection. To explore the mechanism of biofilm formation, mutant strains of Proteus vulgaris XC 2 were generated by Tn5 random transposon insertion. Only one biofilm defective bacterial species was identified from among 500 mutants. Inactivation of the glpC gene coding an anaerobic glycerol-3-phosphate dehydrogenase subunit C was identified by sequence analysis of the biofilm defective strain. Differences were detected in the growth phenotypes of the wild-type and mutant strains under pH, antibiotic, and organic solvent stress conditions. Furthermore, we observed an increase in the phagocytosis of the biofilm defective strain by the mouse macrophage RAW264.7 cell line compared to the wild-type strain. This study shows that the glpC gene plays an important role in biofilm formation, in addition to imparting pH, organic solvent, and antibiotic tolerance, and defense against phagocytosis to Proteus sp. The results further clarified the mechanism of biofilm formation at the genomic level, and indicated the importance of the glpC gene in this process. This data may provide innovative therapeutic measures against P. vulgaris infections; furthermore, as an important crocodile pathogen, this study also has important significance in the protection of Chinese alligators.


Subject(s)
Bacterial Proteins/genetics , Biofilms/growth & development , Glycerolphosphate Dehydrogenase/genetics , Proteus Infections/veterinary , Proteus vulgaris/genetics , Proteus vulgaris/immunology , Adaptation, Physiological/immunology , Alligators and Crocodiles/microbiology , Animals , Bacterial Proteins/immunology , Biofilms/drug effects , Cell Line , Cyclohexanes/pharmacology , DNA Transposable Elements , Escherichia coli/genetics , Escherichia coli/metabolism , Gene Expression , Glycerolphosphate Dehydrogenase/immunology , Hexanes/pharmacology , Hydrogen-Ion Concentration , Immune Evasion , Macrophages/microbiology , Mice , Mutation , Proteus Infections/microbiology , Proteus Infections/pathology , Proteus vulgaris/drug effects , Proteus vulgaris/isolation & purification , Recombinant Proteins/genetics , Recombinant Proteins/immunology
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