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Infection and Immunity, May 2000, p. 2435-2440, Vol. 68, No. 5
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

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

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

Departamento de Microbiología, Facultad de Biología, Universidad de Barcelona, 08071 Barcelona, Spain

Received 12 October 1999/Returned for modification 29 November 1999/Accepted 14 January 2000

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 DH5alpha . 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.

* Corresponding author. Mailing address: Departamento de Microbiología, Facultad de Biología, Universidad de Barcelona, Diagonal 645, 08071 Barcelona, Spain. Phone: 34-934021486. Fax: 34-93-4110592. E-mail: juant{at}porthos.bio.ub.es.

Infection and Immunity, May 2000, p. 2435-2440, Vol. 68, No. 5
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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