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Logo of bmcmicrBioMed Centralsearchsubmit a manuscriptregisterthis articleBMC Microbiology
 
BMC Microbiol. 2012; 12: 201.
Published online Sep 11, 2012. doi:  10.1186/1471-2180-12-201
PMCID: PMC3463446
Novel screening assay for in vivo selection of Klebsiella pneumoniae genes promoting gastrointestinal colonisation
Erik J Boll,1 Lene N Nielsen,1,3 Karen A Krogfelt,1 and Carsten Struvecorresponding author1,2
1Department of Microbiology and Infection Control, Statens Serum Institut, DK-2300, Copenhagen, Denmark
2WHO Collaborating Centre for Reference and Research on Escherichia and Klebsiella, Statens Serum Institut, DK-2300, Copenhagen, Denmark
3Present address: Department of Veterinary Pathobiology, Faculty of Life Sciences, University of Copenhagen, Frederiksberg, Denmark
corresponding authorCorresponding author.
Erik J Boll: ejb/at/ssi.dk; Lene N Nielsen: lenenoerby/at/gmail.com; Karen A Krogfelt: kak/at/ssi.dk; Carsten Struve: cas/at/ssi.dk
Received February 21, 2012; Accepted September 6, 2012.
Abstract
Background
Klebsiella pneumoniae is an important opportunistic pathogen causing pneumonia, sepsis and urinary tract infections. Colonisation of the gastrointestinal (GI) tract is a key step in the development of infections; yet the specific factors important for K. pneumoniae to colonize and reside in the GI tract of the host are largely unknown. To identify K. pneumoniae genes promoting GI colonisation, a novel genomic-library-based approach was employed.
Results
Screening of a K. pneumoniae C3091 genomic library, expressed in E. coli strain EPI100, in a mouse model of GI colonisation led to the positive selection of five clones containing genes promoting persistent colonisation of the mouse GI tract. These included genes encoding the global response regulator ArcA; GalET of the galactose operon; and a cluster of two putative membrane-associated proteins of unknown function. Both ArcA and GalET are known to be involved in metabolic pathways in Klebsiella but may have additional biological actions beneficial to the pathogen. In support of this, GalET was found to confer decreased bile salt sensitivity to EPI100.
Conclusions
The present work establishes the use of genomic-library-based in vivo screening assays as a valuable tool for identification and characterization of virulence factors in K. pneumoniae and other bacterial pathogens.
Keywords: Klebsiella pneumoniae, Genomic library, Mouse model of gastrointestinal colonisation
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