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MOLECULAR KARYOTYPE ANALYSIS AND MAPPING OF HOUSEKEEPING GENES TO CHROMOSOMES OF SELECTED SPECIES COMPLEXES OF LEISHMANIA
Cromossoma
Cariotipagem molecular
PFGE
Eletroforese de gel de campo pulsado
Localização genética
Molecular karyotype
Gene localization
Pulsed-field gel electrophoresis
Leishmania
Affilliation
Universidade do Estado do Rio de Janeiro. Instituto de Biologia. Departamento de Biologia Celular e Genética. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Imunologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Bioquímica Biologia Celular. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Imunologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Bioquímica Biologia Celular. Rio de Janeiro, RJ, Brasil.
Abstract
The molecular karyotypes for 20 reference strains of species complexes of Leishmania were determined by contour-clamped homogeneous electric field (CHEF) electrophoresis. Determination of number/position of chromosome-sized bands and chromosomal DNA locations of housekeeping genes were the two criteria used for differentiating and classifying the Leishmania species. We have established two gel running conditions for optimal separation of chromosomes, which resolved DNA molecules as large as 2,500 kilobase pairs (kb). Chromosomes were polymorphic in number (22-30) and size (200-2,500 kb) of bands among members of five complexes of Leishmania. Although each stock had a distinct karyotype, in general the differences found between strains and/or species within each complex were not clear enough for parasite identification. However, each group showed a specific number of size-concordant DNA molecules, which allowed distinction among the Leishmania complex parasites. Clear differences between the Old and New world groups of parasites or among some New World Leishmania species were also apparent in relation to the chromosome locations of beta-tubulin genes. Based on these results as well as data from other published studies the potential of using DNA karyotype for identifying and classifying leishmanial field isolates is discussed.
Keywords in Portuguese
LeishmaniaCromossoma
Cariotipagem molecular
PFGE
Eletroforese de gel de campo pulsado
Localização genética
Keywords
CromosomeMolecular karyotype
Gene localization
Pulsed-field gel electrophoresis
Leishmania
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