Cloning and molecular analysis of the galE gene of Neisseria meningitidis and its role in lipopolysaccharide biosynthesis.
The galE gene from Haemophilus influenzae was used as a hybridization probe for the galE gene of Neisseria meningitidis Group B, identifying two different homologous loci. Each of the loci was cloned and nucleotide sequence analysis revealed that both loci contained sequences similar to galE. One contained a functional galE gene and mapped to the capsule biosynthetic locus. The second contained only a partial galE-coding sequence, which did not express a functional gene product. A galE mutant meningococcal strain was constructed by transformation with an inactivated galE gene. Analysis of the LPS from the galE mutant strain revealed an apparent reduction in molecular weight and a loss of reactivity with monoclonal antibodies specific for structures known to contain galactose. These results are consistent with an essential role for galE in the incorporation of galactose into meningococcal lipopolysaccharide.
Jennings
M P
MP
Institute of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford, UK.
van der Ley
P
P
Wilks
K E
KE
Maskell
D J
DJ
Poolman
J T
JT
Moxon
E R
ER
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D002238:Carbohydrate Epimerases / Q000235:genetics*; D002240:Carbohydrate Sequence; D003001:Cloning, Molecular; D015342:DNA Probes / Q000235:genetics; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial / Q000235:genetics*; D005816:Genetic Complementation Test; D006193:Haemophilus influenzae / Q000235:genetics; D008070:Lipopolysaccharides / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D005810:Multigene Family; D016296:Mutagenesis; D009345:Neisseria meningitidis / Q000235:genetics* / Q000276:immunology; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012703:Serotyping; D013045:Species Specificity; D014534:UDPglucose 4-Epimerase
D001426:Bacterial Proteins; D015342:DNA Probes; D029968:Escherichia coli Proteins; D008070:Lipopolysaccharides; D002238:Carbohydrate Epimerases; C100033:UDP acetylglucosamine-2-epimerase; C083982:wecB protein, E coli; D014534:UDPglucose 4-Epimerase; C020551:galactose epimerase