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1.
J Med Microbiol ; 56(Pt 8): 1005-1010, 2007 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-17644705

RESUMO

Burkholderia pseudomallei is the causative agent of melioidosis, an infectious disease of humans and animals. Gene clusters which encode capsular polysaccharide (type I O-PS) and LPS (type II O-PS), both of which play roles in virulence, have previously been identified. Here, the identification of two further putative clusters, type III O-PS and type IV O-PS, is reported. Mice challenged with type III O-PS or type IV O-PS mutants showed increased mean times to death (7.8 and 11.6 days) compared to those challenged with wild-type B. pseudomallei (3 days). To investigate the possible roles of polysaccharides in protection, mice were immunized with killed cells of wild-type B. pseudomallei or killed cells of B. pseudomallei with mutations in the O antigen, capsular polysaccharide, type III O-PS or type IV O-PS gene clusters. Immunization with all polysaccharide mutant strains resulted in delayed time to death compared to the naïve controls, following challenge with wild-type B. pseudomallei strain K96243. However, immunization with killed polysaccharide mutant strains conferred different degrees of protection, demonstrating the immunological importance of the polysaccharide clusters on the surface of B. pseudomallei.


Assuntos
Infecções por Burkholderia/microbiologia , Burkholderia pseudomallei/patogenicidade , Ilhas Genômicas/fisiologia , Polissacarídeos Bacterianos/fisiologia , Vacinação , Fatores de Virulência/fisiologia , Animais , Cápsulas Bacterianas/química , Cápsulas Bacterianas/genética , Vacinas Bacterianas/administração & dosagem , Infecções por Burkholderia/prevenção & controle , Burkholderia pseudomallei/genética , Burkholderia pseudomallei/imunologia , Feminino , Genes Bacterianos/genética , Imunização Secundária , Injeções Intraperitoneais , Camundongos , Camundongos Endogâmicos BALB C , Família Multigênica/genética , Antígenos O/genética , Óperon , Mutação Puntual , Virulência
2.
J Microbiol Methods ; 100: 1-7, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24524852

RESUMO

Yersinia pestis, a Gram negative bacterium, causes bubonic and pneumonic plague. Emerging antibiotic resistance in clinical isolates is driving a need to develop novel antibiotics to treat infection by this transmissible and highly virulent pathogen. Proteins required for viability, so called essential genes, are attractive potential therapeutic targets, however, confirmation of essentiality is problematic. For the first time, we report the development of a system that allows the rapid determination of Y. pestis gene essentiality through mutagenesis and inducible expression of a plasmid borne copy of the target gene. Using this approach, we have confirmed the uridine monophosphate kinase PyrH as an essential protein in Y. pestis. This methodology and the tools we have developed will allow the confirmation of other putative essential genes in this dangerous pathogen, and facilitate the identification of novel targets for antimicrobial development.


Assuntos
Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Genes Essenciais , Yersinia pestis/genética , Animais , Modelos Animais de Doenças , Feminino , Expressão Gênica , Técnicas de Inativação de Genes , Camundongos Endogâmicos BALB C , Viabilidade Microbiana , Núcleosídeo-Fosfato Quinase/genética , Peste , Plasmídeos , Virulência , Yersinia pestis/fisiologia
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