ABSTRACT
The molecular karyotype of nine Trypanosoma rangeli strains was analyzed by contour-clamped homogeneous electric field electrophoresis, followed by the chromosomal localization of beta-tubulin, cysteine proteinase, 70 kDa heat shock protein (hsp 70) and actin genes. The T. rangeli strains were isolated from either insects or mammals from El Salvador, Honduras, Venezuela, Colombia, Panama and southern Brazil. Also, T. cruzi CL-Brener clone was included for comparison. Despite the great similarity observed among strains from Brazil, the molecular karyotype of all T. rangeli strains analyzed revealed extensive chromosome polymorphism. In addition, it was possible to distinguish T. rangeli from T. cruzi by the chromosomal DNA electrophoresis pattern. The localization of beta-tubulin genes revealed differences among T. rangeli strains and confirmed the similarity between the isolates from Brazil. Hybridization assays using probes directed to the cysteine proteinase, hsp 70 and actin genes discriminated T. rangeli from T. cruzi, proving that these genes are useful molecular markers for the differential diagnosis between these two species. Numerical analysis based on the molecular karyotype data revealed a high degree of polymorphism among T. rangeli strains isolated from southern Brazil and strains isolated from Central and the northern South America. The T. cruzi reference strain was not clustered with any T. rangeli strain.
Subject(s)
Protozoan Proteins/genetics , Trypanosoma/genetics , Actins/genetics , Animals , Chromosome Mapping , Cysteine Endopeptidases/genetics , Electrophoresis, Gel, Pulsed-Field , Genes, Protozoan/genetics , Genetic Variation , HSP70 Heat-Shock Proteins/genetics , Latin America , Trypanosoma/enzymology , Trypanosoma/isolation & purification , Tubulin/geneticsABSTRACT
The molecular karyotype of nine Trypanosoma rangeli strains was analyzed by contour-clamped homogeneous electric field electrophoresis, followed by the chromosomal localization of ß-tubulin, cysteine proteinase, 70 kDa heat shock protein (hsp 70) and actin genes. The T. rangeli strains were isolated from either insects or mammals from El Salvador, Honduras, Venezuela, Colombia, Panama and southern Brazil. Also, T. cruzi CL-Brener clone was included for comparison. Despite the great similarity observed among strains from Brazil, the molecular karyotype of all T. rangeli strains analyzed revealed extensive chromosome polymorphism. In addition, it was possible to distinguish T. rangeli from T. cruzi by the chromosomal DNA electrophoresis pattern. The localization of ß-tubulin genes revealed differences among T. rangeli strains and confirmed the similarity between the isolates from Brazil. Hybridization assays using probes directed to the cysteine proteinase, hsp 70 and actin genes discriminated T. rangeli from T. cruzi, proving that these genes are useful molecular markers for the differential diagnosis between these two species. Numerical analysis based on the molecular karyotype data revealed a high degree of polymorphism among T. rangeli strains isolated from southern Brazil and strains isolated from Central and the northern South America. The T. cruzi reference strain was not clustered with any T. rangeli strain
Subject(s)
Animals , Actins/genetics , Chromosome Mapping , Cysteine Endopeptidases/genetics , HSP70 Heat-Shock Proteins/genetics , Protozoan Proteins/genetics , Trypanosoma/genetics , Brazil , Colombia , El Salvador , Electrophoresis, Gel, Pulsed-Field , Genes, Protozoan/genetics , Genetic Variation , Honduras , Karyotyping , Panama , Protozoan Proteins/genetics , Trypanosoma/enzymology , Trypanosoma/isolation & purification , Tubulin/genetics , VenezuelaABSTRACT
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.
Subject(s)
Chromosome Mapping , Genes, Protozoan , Karyotyping , Leishmania/genetics , Animals , DNA, Protozoan/genetics , Electrophoresis, Gel, Pulsed-Field/methods , Leishmania/classification , Species SpecificityABSTRACT
Elevation of the NaCl concentration in the growth medium of L-A9 cells caused an inhibition of the protein synthesis accompanied by a complete breakdown of polyribosomes. However, a complete recovery of the rate of protein synthesis was observed when isotonicity was restored. In Marituba virus infected cells, protein synthesis became resistant to hypertonic treatment. Under hypertonic conditions cellular protein synthesis was selectively suppressed and an enhancement of virus proteins was observed. Analysis of the virus specific proteins by polyacrylamide gel electrophoresis revealed that the synthesis of G1 was unalterable, and N was stimulated.
