1887

Abstract

Members of the complex are Gram-negative β-proteobacteria that are classified into nine genomic species or genomovars. Some representatives of this group of bacteria, such as (genomovar II) and (genomovar III), are considered to be dangerous pathogens for cystic fibrosis (CF) patients because of their capacity to colonize CF lungs. The gene, which encodes the peptidoglycan-associated outer-membrane lipoprotein (PAL), was detected in the genome of sp. LB 400 by a similarity search that was based on the sequence of the PAL, OprL. Primers that could amplify part of from LMG 13010 were designed. This PCR fragment was used as a probe for screening of a genomic bank, allowing cloning of the complete gene. The complete gene could be PCR-amplified from DNA from all genomovars. The sequences of these genes showed a high degree of conservation (> 95 %) among different species of the complex. OpcL protein that was purified from LMG 13010 was used to generate mouse polyclonal antisera against OpcL. The OpcL protein could be produced in and detected in outer-membrane fractions by Western blot. cells were labelled by immunofluorescence staining using antibodies against OpcL, but only after treatment with EDTA and SDS. The gene could be amplified directly from the sputa of 15 CF patients who were known to be colonized by ; sequence data derived from the amplicons identified the colonizing strains as (genomovar III, = 14) and ( = 1).

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2004-05-01
2019-11-14
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