This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Que, Y.-A.
Right arrow Articles by Moreillon, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Que, Y.-A.
Right arrow Articles by Moreillon, P.

 Previous Article  |  Next Article 

Infection and Immunity, June 2000, p. 3516-3522, Vol. 68, No. 6
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Expression of Staphylococcus aureus Clumping Factor A in Lactococcus lactis subsp. cremoris Using a New Shuttle Vector

Yok-Ai Que,1 Jacques-Antoine Haefliger,2 Patrick Francioli,1 and Philippe Moreillon1,*

Division of Infectious Diseases,1 Laboratory of Molecular Biology,2 Department of Internal Medicine, Centre Hospitalier Universitaire Vaudois, 1011 Lausanne, Switzerland

Received 22 November 1999/Returned for modification 14 January 2000/Accepted 24 February 2000

Staphylococcus aureus harbors redundant adhesins mediating tissue colonization and infection. To evaluate their intrinsic role outside of the staphylococcal background, a system was designed to express them in Lactococcus lactis subsp. cremoris 1363. This bacterium is devoid of virulence factors and has a known genetic background. A new Escherichia coli-L. lactis shuttle and expression vector was constructed for this purpose. First, the high-copy-number lactococcal plasmid pIL253 was equipped with the oriColE1 origin, generating pOri253 that could replicate in E. coli. Second, the lactococcal promoters P23 or P59 were inserted at one end of the pOri253 multicloning site. Gene expression was assessed by a luciferase reporter system. The plasmid carrying P23 (named pOri23) expressed luciferase constitutively at a level 10,000 times greater than did the P59-containing plasmid. Transcription was absent in E. coli. The staphylococcal clumping factor A (clfA) gene was cloned into pOri23 and used as a model system. Lactococci carrying pOri23-clfA produced an unaltered and functional 130-kDa ClfA protein attached to their cell walls. This was indicated both by the presence of the protein in Western blots of solubilized cell walls and by the ability of ClfA-positive lactococci to clump in the presence of plasma. ClfA-positive lactococci had clumping titers (titer of 4,112) similar to those of S. aureus Newman in soluble fibrinogen and bound equally well to solid-phase fibrinogen. These experiments provide a new way to study individual staphylococcal pathogenic factors and might complement both classical knockout mutagenesis and modern in vivo expression technology and signature tag mutagenesis.

* Corresponding author. Mailing address: Division of Infectious Diseases, Department of Internal Medicine, Centre Hospitalier Universitaire Vaudois, 1011 Lausanne, Switzerland. Phone: 41-21-314-10-26. Fax: 41-21-314-10-36. E-mail: pmoreill{at}hola.hospvd.ch.

Infection and Immunity, June 2000, p. 3516-3522, Vol. 68, No. 6
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Innocentin, S., Guimaraes, V., Miyoshi, A., Azevedo, V., Langella, P., Chatel, J.-M., Lefevre, F. (2009). Lactococcus lactis Expressing either Staphylococcus aureus Fibronectin-Binding Protein A or Listeria monocytogenes Internalin A Can Efficiently Internalize and Deliver DNA in Human Epithelial Cells. Appl. Environ. Microbiol. 75: 4870-4878 [Abstract] [Full Text]  
  • Zhang, J., Biswas, I. (2009). 3'-Phosphoadenosine-5'-Phosphate Phosphatase Activity Is Required for Superoxide Stress Tolerance in Streptococcus mutans. J. Bacteriol. 191: 4330-4340 [Abstract] [Full Text]  
  • Ma, Y., Bryant, A. E., Salmi, D. B., McIndoo, E., Stevens, D. L. (2009). vfr, a Novel Locus Affecting Cysteine Protease Production in Streptococcus pyogenes. J. Bacteriol. 191: 3189-3194 [Abstract] [Full Text]  
  • Biswas, I., Jha, J. K., Fromm, N. (2008). Shuttle expression plasmids for genetic studies in Streptococcus mutans. Microbiology 154: 2275-2282 [Abstract] [Full Text]  
  • Piroth, L., Que, Y.-A., Widmer, E., Panchaud, A., Piu, S., Entenza, J. M., Moreillon, P. (2008). The Fibrinogen- and Fibronectin-Binding Domains of Staphylococcus aureus Fibronectin-Binding Protein A Synergistically Promote Endothelial Invasion and Experimental Endocarditis. Infect. Immun. 76: 3824-3831 [Abstract] [Full Text]  
  • Kerrigan, S. W., Clarke, N., Loughman, A., Meade, G., Foster, T. J., Cox, D. (2008). Molecular Basis for Staphylococcus aureus-Mediated Platelet Aggregate Formation Under Arterial Shear In Vitro. Arterioscler. Thromb. Vasc. Bio. 28: 335-340 [Abstract] [Full Text]  
  • Arsene, S., Leclercq, R. (2007). Role of a qnr-Like Gene in the Intrinsic Resistance of Enterococcus faecalis to Fluoroquinolones. Antimicrob. Agents Chemother. 51: 3254-3258 [Abstract] [Full Text]  
  • Arrecubieta, C., Lee, M.-H., Macey, A., Foster, T. J., Lowy, F. D. (2007). SdrF, a Staphylococcus epidermidis Surface Protein, Binds Type I Collagen. J. Biol. Chem. 282: 18767-18776 [Abstract] [Full Text]  
  • Barnett, T. C., Bugrysheva, J. V., Scott, J. R. (2007). Role of mRNA Stability in Growth Phase Regulation of Gene Expression in the Group A Streptococcus. J. Bacteriol. 189: 1866-1873 [Abstract] [Full Text]  
  • Baums, C. G., Kaim, U., Fulde, M., Ramachandran, G., Goethe, R., Valentin-Weigand, P. (2006). Identification of a Novel Virulence Determinant with Serum Opacification Activity in Streptococcus suis.. Infect. Immun. 74: 6154-6162 [Abstract] [Full Text]  
  • Biswas, S., Biswas, I. (2006). Regulation of the Glucosyltransferase (gtfBC) Operon by CovR in Streptococcus mutans. J. Bacteriol. 188: 988-998 [Abstract] [Full Text]  
  • Dalton, T. L., Collins, J. T., Barnett, T. C., Scott, J. R. (2006). RscA, a Member of the MDR1 Family of Transporters, Is Repressed by CovR and Required for Growth of Streptococcus pyogenes under Heat Stress. J. Bacteriol. 188: 77-85 [Abstract] [Full Text]  
  • Domanski, P. J., Patel, P. R., Bayer, A. S., Zhang, L., Hall, A. E., Syribeys, P. J., Gorovits, E. L., Bryant, D., Vernachio, J. H., Hutchins, J. T., Patti, J. M. (2005). Characterization of a Humanized Monoclonal Antibody Recognizing Clumping Factor A Expressed by Staphylococcus aureus. Infect. Immun. 73: 5229-5232 [Abstract] [Full Text]  
  • Que, Y.-A., Haefliger, J.-A., Piroth, L., Francois, P., Widmer, E., Entenza, J. M., Sinha, B., Herrmann, M., Francioli, P., Vaudaux, P., Moreillon, P. (2005). Fibrinogen and fibronectin binding cooperate for valve infection and invasion in Staphylococcus aureus experimental endocarditis. JEM 201: 1627-1635 [Abstract] [Full Text]  
  • Prunier, A.-L., Leclercq, R. (2005). Role of mutS and mutL Genes in Hypermutability and Recombination in Staphylococcus aureus. J. Bacteriol. 187: 3455-3464 [Abstract] [Full Text]  
  • Reilley, S., Wenzel, E., Reynolds, L., Bennett, B., Patti, J. M., Hetherington, S. (2005). Open-Label, Dose Escalation Study of the Safety and Pharmacokinetic Profile of Tefibazumab in Healthy Volunteers. Antimicrob. Agents Chemother. 49: 959-962 [Abstract] [Full Text]  
  • Pietrocola, G., Schubert, A., Visai, L., Torti, M., Fitzgerald, J. R., Foster, T. J., Reinscheid, D. J., Speziale, P. (2005). FbsA, a fibrinogen-binding protein from Streptococcus agalactiae, mediates platelet aggregation. Blood 105: 1052-1059 [Abstract] [Full Text]  
  • Schubert, A., Zakikhany, K., Pietrocola, G., Meinke, A., Speziale, P., Eikmanns, B. J., Reinscheid, D. J. (2004). The Fibrinogen Receptor FbsA Promotes Adherence of Streptococcus agalactiae to Human Epithelial Cells. Infect. Immun. 72: 6197-6205 [Abstract] [Full Text]  
  • Dalton, T. L., Scott, J. R. (2004). CovS Inactivates CovR and Is Required for Growth under Conditions of General Stress in Streptococcus pyogenes. J. Bacteriol. 186: 3928-3937 [Abstract] [Full Text]  
  • Hall, A. E., Domanski, P. J., Patel, P. R., Vernachio, J. H., Syribeys, P. J., Gorovits, E. L., Johnson, M. A., Ross, J. M., Hutchins, J. T., Patti, J. M. (2003). Characterization of a Protective Monoclonal Antibody Recognizing Staphylococcus aureus MSCRAMM Protein Clumping Factor A. Infect. Immun. 71: 6864-6870 [Abstract] [Full Text]  
  • Bates, C. S., Montanez, G. E., Woods, C. R., Vincent, R. M., Eichenbaum, Z. (2003). Identification and Characterization of a Streptococcus pyogenes Operon Involved in Binding of Hemoproteins and Acquisition of Iron. Infect. Immun. 71: 1042-1055 [Abstract] [Full Text]  
  • Wolz, C., Goerke, C., Landmann, R., Zimmerli, W., Fluckiger, U. (2002). Transcription of Clumping Factor A in Attached and Unattached Staphylococcus aureus In Vitro and during Device-Related Infection. Infect. Immun. 70: 2758-2762 [Abstract] [Full Text]  
  • Ribeiro, L. A., Azevedo, V., Le Loir, Y., Oliveira, S. C., Dieye, Y., Piard, J.-C., Gruss, A., Langella, P. (2002). Production and Targeting of the Brucella abortus Antigen L7/L12 in Lactococcus lactis: a First Step towards Food-Grade Live Vaccines against Brucellosis. Appl. Environ. Microbiol. 68: 910-916 [Abstract] [Full Text]  
  • Que, Y.-A., Francois, P., Haefliger, J.-A., Entenza, J.-M., Vaudaux, P., Moreillon, P. (2001). Reassessing the Role of Staphylococcus aureus Clumping Factor and Fibronectin-Binding Protein by Expression in Lactococcus lactis. Infect. Immun. 69: 6296-6302 [Abstract] [Full Text]  
  • Hartford, O., O'Brien, L., Schofield, K., Wells, J., Foster, T. J. (2001). The Fbe (SdrG) protein of Staphylococcus epidermidis HB promotes bacterial adherence to fibrinogen. Microbiology 147: 2545-2552 [Abstract] [Full Text]  
  • Stutzmann Meier, P., Entenza, J. M., Vaudaux, P., Francioli, P., Glauser, M. P., Moreillon, P. (2001). Study of Staphylococcus aureus Pathogenic Genes by Transfer and Expression in the Less Virulent Organism Streptococcus gordonii. Infect. Immun. 69: 657-664 [Abstract] [Full Text]  
  • Sinha, B., Francois, P., Que, Y.-A., Hussain, M., Heilmann, C., Moreillon, P., Lew, D., Krause, K.-H., Peters, G., Herrmann, M. (2000). Heterologously Expressed Staphylococcus aureus Fibronectin-Binding Proteins Are Sufficient for Invasion of Host Cells. Infect. Immun. 68: 6871-6878 [Abstract] [Full Text]  
  • Entenza, J. M., Foster, T. J., Ni Eidhin, D., Vaudaux, P., Francioli, P., Moreillon, P. (2000). Contribution of Clumping Factor B to Pathogenesis of Experimental Endocarditis due to Staphylococcus aureus. Infect. Immun. 68: 5443-5446 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»