Use of the complete genome sequence information of Haemophilus influenzae strain Rd to investigate lipopolysaccharide biosynthesis.
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.
Hood
D W
DW
University of Oxford, Department of Paediatrics, John Radcliffe Hospital, Headington, UK. dhood@worf.molbiol.ox.ac.uk
Deadman
M E
ME
Allen
T
T
Masoud
H
H
Martin
A
A
Brisson
J R
JR
Fleischmann
R
R
Venter
J C
JC
Richards
J C
JC
Moxon
E R
ER
D000818:Animals; D000831:Animals, Newborn; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D016680:Genome, Bacterial; D005998:Glycine / Q000031:analogs & derivatives / Q000737:chemistry; D006193:Haemophilus influenzae / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D015151:Immunoblotting; D008070:Lipopolysaccharides / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D013058:Mass Spectrometry; D008958:Models, Molecular; D009010:Monte Carlo Method; D016296:Mutagenesis; D051381:Rats; D014774:Virulence
D008070:Lipopolysaccharides; D005998:Glycine; C100184:tricine