Identification and characterization of the N-acetylglucosamine glycosyltransferase gene of Haemophilus ducreyi.
Haemophilus ducreyi is the causative agent of chancroid, a sexually transmitted ulcerative disease. In the present study, the Neisseria gonorrhoeae lgtA lipooligosaccharide glycosyltransferase gene was used to identify a homologue in the genome of H. ducreyi. The putative H. ducreyi glycosyltransferase gene (designated lgtA) was cloned and insertionally inactivated, and an isogenic mutant was constructed. Structural studies demonstrated that the lipooligosaccharide isolated from the mutant strain lacked N-acetylglucosamine and distal sugars found in the lipooligosaccharide produced by the parental strain. The isogenic mutant was transformed with a recombinant plasmid containing the putative glycosyltransferase gene. This strain produced the lipooligosaccharide glycoforms produced by the parental strain, confirming that the lgtA gene encodes the N-acetylglucosamine glycosyltransferase.
Sun
Shuhua
S
Columbus Children's Research Institute, Department of Pediatrics, The Ohio State University, Columbus, Ohio 43205, USA.
Scheffler
N Karoline
NK
Gibson
Bradford W
BW
Wang
Jing
J
Munson
Robert S
RS
D001426:Bacterial Proteins; D001483:Base Sequence; D002240:Carbohydrate Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D006191:Haemophilus ducreyi / Q000201:enzymology* / Q000235:genetics*; D006801:Humans; D008070:Lipopolysaccharides / Q000096:biosynthesis / Q000737:chemistry; D008969:Molecular Sequence Data; D009154:Mutation; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D008070:Lipopolysaccharides; C023023:lipid-linked oligosaccharides; C401409:LgtA protein, bacteria; D017351:N-Acetylglucosaminyltransferases