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Cloning and Sequencing of the Klebsiella pneumoniae O5 wb Gene Cluster and Its Role in Pathogenesis

By Susana Merino, Maria Altarriba, Luis Izquierdo, María Mercé Nogueras, Miguel Regué and Juan M. Tomás

Abstract

One representative recombinant clone encoding Klebsiella pneumoniae O5-antigen lipopolysaccharide (LPS) was found upon screening for serum resistance in a cosmid-based genomic library of K. pneumoniae KT769 (O5:K57) introduced into Escherichia coli DH5α. A total of eight open reading frames (wbO5 gene cluster) were necessary to produce K. pneumoniae O5-antigen LPS in E. coli K-12. The enzymatic activities proposed for the wbO5 gene cluster are in agreement with the activities proposed for the biosynthesis of K. pneumoniae O5-antigen LPS. Using the complete DNA sequence of the K. pneumoniae wbO5 gene cluster, we obtained (by single or double recombination) genetically well-characterized mutants devoid only of this O5-antigen LPS. Finally, using these O5− mutants and the corresponding wild-type strains or complemented mutants with the wbO5 gene cluster (O5+ strains), we found that the presence of K. pneumoniae O5-antigen LPS is essential for some pathogenic features like serum resistance, adhesion to uroepithelial cells, and colonization (experimental infections) of the urinary tract in rats

Topics: Molecular and Cellular Pathogenesis
Publisher: American Society for Microbiology
Year: 2000
OAI identifier: oai:pubmedcentral.nih.gov:97443
Provided by: PubMed Central
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