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 Vilela, R.
Right arrow Articles by de Resende, M. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Vilela, R.
Right arrow Articles by de Resende, M. A.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, August 2005, p. 3657-3661, Vol. 43, No. 8
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.8.3657-3661.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Molecular Model for Studying the Uncultivated Fungal Pathogen Lacazia loboi

Raquel Vilela,1 Leonel Mendoza,2* Patricia S. Rosa,3 Andréa Faria Fernandes Belone,3 Suzana Madeira,3 Diltor Vladimir Araújo Opromolla,3 and Maria Aparecida de Resende1

Department of Microbiology, Universidade Federal de Minas Gerais, Minas Gerais, Brazil,1 Medical Technology Program, Microbiology and Molecular Genetics, Michigan State University, East Lansing, Michigan,2 Instituto Lauro De Souza Lima, Bauru, São Paulo, Brazil3

Received 2 March 2005/ Returned for modification 11 March 2005/ Accepted 13 April 2005

Lacazia loboi is an uncultivated fungal pathogen of humans and dolphins that causes cutaneous and subcutaneous infections only in the tropical areas of the Americas. It was recently found by phylogenetic analysis that this unusual pathogen is closely related to Paracoccidioides brasiliensis and to the other fungal dimorphic members of the order Onygenales. That original phylogenetic study used universal primers to amplify well-known genes. However, this approach cannot be applied to the study of other proteins. We have developed a strategy for studying the gene encoding the gp43 homologous protein of P. brasiliensis in L. loboi. The gp43 protein was selected because it has been found that this P. brasiliensis antigen strongly reacts when it is used to test sera from patients with lacaziosis. The principle behind this idea was to obtain the gp43 amino acid sequence of P. brasiliensis and other homologous fungal sequences from GenBank and design primers from their aligned conserved regions. These sets of primers were used to amplify the selected regions with genomic DNA extracted from the yeast-like cells of L. loboi from experimentally infected mice. Using this approach, we amplified 483 bp of the L. loboi gp43-like gene. These sequences had 85% identity at the nucleotide level and 75% identity with the deduced amino acid sequences of the P. brasiliensis gp43 protein. The identity of the 483-bp DNA fragment was confirmed by phylogenetic analysis. This analysis revealed that the L. loboi gp43-like deduced amino acid sequence formed a strongly supported (100%) sister group with several P. brasiliensis gp43 sequences and that this taxon in turn was linked to the other fungal sequences used in this analysis. This study shows that the use of a molecular model for investigation of the genes encoding important proteins in L. loboi is feasible.

* Corresponding author. Mailing address: Medical Technology Program, Michigan State University, 322 North Kedzie Hall, East Lansing, MI 48824-1031. Phone: (517) 353-7800. Fax: (517) 432-2006. E-mail: mendoza9{at}msu.edu.

Journal of Clinical Microbiology, August 2005, p. 3657-3661, Vol. 43, No. 8
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.8.3657-3661.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Mendoza, L., Belone, A. F. F., Vilela, R., Rehtanz, M., Bossart, G. D., Reif, J. S., Fair, P. A., Durden, W. N., St. Leger, J., Travassos, L. R., Rosa, P. S. (2008). Use of Sera from Humans and Dolphins with Lacaziosis and Sera from Experimentally Infected Mice for Western Blot Analyses of Lacazia loboi Antigens. CVI 15: 164-167 [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»