Cloning, sequencing, and characterization of the lipopolysaccharide biosynthetic enzyme heptosyltransferase I gene (waaC) from Campylobacter jejuni and Campylobacter coli.
Campylobacter jejuni and Campylobacter coli are common causes of gastrointestinal disease and a proportion of C. jejuni infections have been shown to be associated with the Guillain-Barré syndrome. The waaC gene from Campylobacter coli, involved in lipopolysaccharide core biosynthesis, was cloned by complementation of a heptose-deficient strain of Salmonella typhimurium, as judged by novobiocin sensitivity, lipopolysaccharide (LPS)-specific phage sensitivity, and polyacrylamide-resolved lipopolysaccharide profiles. The C. jejuni waaC gene was subsequently cloned using the waaC gene isolated from C. coli as a probe. The C. jejuni and C. coli waaC genes are capable of encoding proteins of 342 amino acids with calculated molecular masses of 39381Da and 39317Da, respectively. Sequence and in-vitro analyses suggested that the C. coli waaC gene may be transcribed from its own promoter. Translation of the C. coli waaC gene in a cell-free system yielded a protein with a Mr of 39000. The waaC gene was detected in every C. jejuni and C. coli isolate tested as judged by dot-blot hybridization analysis. Southern hybridization analysis indicated that both Campylobacter species contain a single copy of the waaC gene. Unlike Escherichia coli and S. typhimurium isolates, the waaC gene in C. jejuni and C. coli isolates does not appear to be linked to the waaF (rfaF) gene.
Klena
J D
JD
Department of Plant and Microbial Science, University of Canterbury, Christchurch, New Zealand.
Gray
S A
SA
Konkel
M E
ME
D001483:Base Sequence; D017000:Campylobacter coli / Q000201:enzymology* / Q000235:genetics* / Q000472:pathogenicity; D016123:Campylobacter jejuni / Q000201:enzymology* / Q000235:genetics* / Q000472:pathogenicity; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D016695:Glycosyltransferases / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008070:Lipopolysaccharides / Q000096:biosynthesis; D008969:Molecular Sequence Data; D020161:Physical Chromosome Mapping; D016133:Polymerase Chain Reaction; D012486:Salmonella typhimurium / Q000201:enzymology / Q000235:genetics; D013045:Species Specificity; D014774:Virulence / Q000235:genetics
D017931:DNA Primers; D004269:DNA, Bacterial; D008070:Lipopolysaccharides; D016695:Glycosyltransferases; C080934:lipooligosaccharide 1,5-heptosyltransferase