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Journal of Clinical Microbiology, March 1998, p. 742-747, Vol. 36, No. 3
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Electrophoretic Karyotypes and Genome Sizing of the Pathogenic Fungus Paracoccidioides brasiliensis

Maria Isabel Nogueira Cano,1 Patrícia Silva Cisalpino,2 Ivan Galindo,3 José Luiz Ramírez,3 Renato Arruda Mortara,1 and José Franco da Silveira1,*

Department of Microbiology, Immunology, and Parasitology, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo SP,1 and Department of Microbiology, Universidade Federal de Minas Gerais, Belo Horizonte MG,2 Brazil, and Centro de Biologia Molecular, Universidad Central de Venezuela, Caracas, Venezuela3

Received 13 August 1997/Returned for modification 13 September 1997/Accepted 22 November 1997

Here we present the karyotype analysis and genome sizing of Paracoccidioides brasiliensis, a pathogen refractory to conventional genetic analysis. We have established pulsed-field gel electrophoresis (PFGE) conditions to resolve the high-molecular-weight chromosomal bands of two clinical isolates of P. brasiliensis. Both isolates showed four megabase-sized bands, ranging from 2.0 to 10.0 Mbp. Significant differences in chromosome sizes and in the chromosomal location of genes for the gp43 antigen and chitin synthase were found. Different technical approaches were employed to estimate the DNA content and to define the ploidy of P. brasiliensis. An estimated genome size in the range of 45.7 to 60.9 Mbp was provided by the analysis of data generated by measuring the amplitude of fluorescence intensity of DAPI (4',6-diamidino-2-phenylindole)-stained nuclei (by confocal microscopy). The nuclear genome size estimated by confocal microscopy is twice that estimated by the average sum of the molecular weight of chromosome-sized DNA molecules by PFGE, suggesting that each separated P. brasiliensis chromosomal band is diploid.

* Corresponding author. Mailing address: Department of Microbiology, Immunology, and Parasitology, Escola Paulista de Medicina, Universidade Federal de São Paulo, Rua Botucatu 862, CEP 04023-062 São Paulo SP, Brazil. Phone: (55 11) 576 4532. Fax: (55 11) 571 1095. E-mail: franco.dmip{at}epm.br.

Journal of Clinical Microbiology, March 1998, p. 742-747, Vol. 36, No. 3
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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