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Infection and Immunity, October 2002, p. 5887-5892, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5887-5892.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification and Characterization of the N-Acetylglucosamine Glycosyltransferase Gene of Haemophilus ducreyi

Shuhua Sun,^1,2 N. Karoline Scheffler,³ Bradford W. Gibson,^3,4 Jing Wang,¹ and Robert S. Munson Jr.^1,2,5*

Columbus Children's Research Institute,¹ Department of Pediatrics,⁵ Department of Molecular Virology, Immunology, and Medical Genetics, The Ohio State University, Columbus, Ohio 43205,² Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94143,³ Buck Institute for Age Research, Novato, California 94945⁴

Received 12 April 2002/ Returned for modification 13 May 2002/ Accepted 3 July 2002

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.

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* Corresponding author. Mailing address: Columbus Children's Research Institute, Rm. W402, 700 Children's Dr., Columbus, OH 43205. Phone: (614) 722-2680. Fax: (614) 722-3273. E-mail: munsonr{at}pediatrics.ohio-state.edu.

Editor: B. B. Finlay

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Infection and Immunity, October 2002, p. 5887-5892, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5887-5892.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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