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Journal of Bacteriology, August 2003, p. 4393-4401, Vol. 185, No. 15
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.15.4393-4401.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Molecular Analysis of the Enterococcus faecalis Serotype 2 Polysaccharide Determinant

Lynn E. Hancock,1 Brett D. Shepard,1 and Michael S. Gilmore1,2*

Department of Microbiology and Immunology,1 Department of Ophthalmology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 731042

Received 2 December 2002/ Accepted 25 April 2003

We previously described a 15-kb genetic cluster consisting of 11 open reading frames (cps2A to cps2K) of Enterococcus faecalis FA2-2 that is responsible for the production of the serotype 2 capsular polysaccharide. By using transcriptional fusions to a promoterless lacZ gene, we identified two independent promoters related to the expression of the polysaccharide. Both transcription initiation sites were mapped by primer extension. Reverse transcription-PCR (RT-PCR) demonstrated the transcriptional linkage of genes present in both transcripts. Real-time RT-PCR quantification of transcripts revealed maximum transcription during log phase growth, an observation confirmed by promoter fusion studies. The heterologous expression of this pathway in Escherichia coli caused reactivity with E. faecalis type 2 antiserum, thus demonstrating the essential role of this pathway in the synthesis of the type-specific polysaccharide.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Biomedical Research Center, Rm. 354A, Oklahoma City, OK 73104. Phone: (405) 271-1863. Fax: (405) 271-8128. E-mail: michael-gilmore{at}ouhsc.edu.

Journal of Bacteriology, August 2003, p. 4393-4401, Vol. 185, No. 15
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.15.4393-4401.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Thurlow, L. R., Thomas, V. C., Hancock, L. E. (2009). Capsular Polysaccharide Production in Enterococcus faecalis and Contribution of CpsF to Capsule Serospecificity. J. Bacteriol. 191: 6203-6210 [Abstract] [Full Text]  
  • Bourgogne, A., Singh, K. V., Fox, K. A., Pflughoeft, K. J., Murray, B. E., Garsin, D. A. (2007). EbpR Is Important for Biofilm Formation by Activating Expression of the Endocarditis and Biofilm-Associated Pilus Operon (ebpABC) of Enterococcus faecalis OG1RF. J. Bacteriol. 189: 6490-6493 [Abstract] [Full Text]  
  • Hufnagel, M., Hancock, L. E., Koch, S., Theilacker, C., Gilmore, M. S., Huebner, J. (2004). Serological and Genetic Diversity of Capsular Polysaccharides in Enterococcus faecalis. J. Clin. Microbiol. 42: 2548-2557 [Abstract] [Full Text]  


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