Use of the complete genome sequence information of Haemophilus influenzae strain Rd to investigate lipopolysaccharide biosynthesis

Authors: Hood, Derek W.1; Deadman, Mary E.1; Allen, Tina1; Masoud, Hussein2; Martin, Adele2; Brisson, Jean Robert2; Fleischmann, Robert3; Venter, J. Craig3; Richards, James C.2; Moxon, E. Richard1

Source: Molecular Microbiology, Volume 22, Number 5, December 1996 , pp. 951-965(15)

Publisher: Blackwell Publishing

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Abstract:

The availability of the complete 1.83-megabase-pair sequence of the Haemophilus influenzae strain Rd genome has facilitated significant progress in investigating the biology of H. influenzae lipopolysaccharide (LPS), a major virulence determinant of this human pathogen. By searching the H. influenzae genomic database, with sequences of known LPS biosynthetic genes from other organisms, we identified and then cloned 25 candidate LPS genes. Construction of mutant strains and characterization of the LPS by reactivity with monoclonal antibodies, PAGE fractionation patterns and electrospray mass spectrometry comparative analysis have confirmed a potential role in LPS biosynthesis for the majority of these candidate genes. Virulence studies in the infant rat have allowed us to estimate the minimal LPS structure required for intravascular dissemination. This study is one of the first to demonstrate the rapidity, economy and completeness with which novel biological information can be accessed once the complete genome sequence of an organism is available.

Document Type: Research article

DOI: 10.1046/j.1365-2958.1996.01545.x

Affiliations: 1: Molecular Infectious Diseases Group, University of Oxford Department of Paediatrics, Institute of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford, OX3 9DU, UK. 2: Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, Canada, K1A 0R6. 3: The Institute for Genomic Research, 9712 Medical Centre Drive, Rockville, Maryland 20850, USA.

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