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Infection and Immunity, February 2000, p. 815-823, Vol. 68, No. 2
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Analysis of a Gene Cluster of Enterococcus faecalis Involved in Polysaccharide Biosynthesis

Yi Xu,1,2,dagger Kavindra V. Singh,2,3 Xiang Qin,2,3 Barbara E. Murray,2,3 and George M. Weinstock1,2,*

Department of Microbiology and Molecular Genetics,1 Division of Infectious Diseases, Department of Medicine,3 and Center for the Study of Emerging and Re-emerging Pathogens,2 University of Texas Medical School, Houston, Texas 77030

Received 22 July 1999/Returned for modification 8 September 1999/Accepted 9 November 1999

Previously, we described a gene cluster of Enterococcus faecalis OG1RF that produced an antigenic polysaccharide when cloned in Escherichia coli. The polysaccharide antigen was not detectable in E. faecalis strains, however. Here, we show by reverse transcriptase-PCR that the 16 genes in this region are transcribed in OG1RF. Gene disruption of orfde4, encoding a putative glycosyl transferase, and orfde6, a putative dTDP-rhamnose biosynthesis gene, generated two OG1RF mutants. The mutants showed delayed killing and a higher 50% lethal dose in a mouse peritonitis model. In addition, two mucoid E. faecalis isolates from patients with chronic urinary tract infections were found to produce the polysaccharide antigen.

* Corresponding author. Mailing address: Department of Microbiology and Molecular Genetics, University of Texas Medical School, 6431 Fannin St., Houston, TX 77030. Phone: (713) 500-6083. Fax: (713) 500-5499. E-mail: georgew{at}utmmg.med.uth.tmc.edu.

dagger Present address: Center for Extracellular Matrix Biology, Albert B. Alkek Institute of Biosciences and Technology, Texas A&M University, Houston, TX 77030-3303.

Infection and Immunity, February 2000, p. 815-823, Vol. 68, No. 2
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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