Molecular cloning of two Pseudomonas flagellin genes and basal body structural genes

摘要: Pseudomonas putida strains PaW8 and PRS2000 produce flagellins with apparent molecular masses of 81 kDa 50 respectively. Two Tn5 insertion mutants P. lacking the ability to bind flagellin-specific monoclonal antibody MLV1 were isolated. Mutant PaW8-flg2 contained a within 2.6 kb EcoRI fragment chromosome carrying putative basal body genes. DNA deduced protein sequences suggested presence on this two complete genes homologous flgH flgl from Salmonella typhimurium. The occurred in locus appeared severely reduce expression flagellin gene. A Tn5-containing second mutant, PaW8-flg1, was cloned found contain that hybridized strongly aeruginosa 2.3 HindIII containing all but 62 bp gene used as probe identify clones equivalent PRS2000. Flagellin both sequenced their amino acid deduced. Both conserved amino- carboxy-terminal regions when compared other flagellins, central region being more variable. epitope for is likely lie flagellin. mass (68 kDa) differed significantly its estimated by PAGE, possibly consequence post-translational modification. This not case PRS2000, where predicted similar.