ABSTRACT
Many protozoan parasites represent an important group of human pathogens. Pulsed Field Gradient Gel Electrophoresis (PFGE) analysis has been an important tool fundamental genetic studies of parasites like Trypanosoma, Leishmania Giardia or the human malaria Plasmodium falciparum. We present PFGE conditions allowing a high resolution separation of chromosomes ranging from 500 to 4000 kb within a two day electrophoresis run. In addition, we present conditions for separating large chromosomes (2000-6000 kb) within 36 hr. We demontrate that the application of two dimentional PFGE (2D-PFGE) technique to parasite karyotypes is a very useful method for the analysis of dispersed gene families and comparative studies of the intrachomosomal genome organization.
Subject(s)
Animals , Chromosome Mapping , Chromosomes , Eukaryota/genetics , Electrophoresis, Gel, Pulsed-Field , KaryotypingABSTRACT
Our research work group has been interested in the study of the ribosomal RNA and 5S gene systems from Trypanosoma cruzi. Our contributions span from the general description of a multifragmented molecular system, to the sequence analysis of some ribosomal RNA coding regions. From the latter, we have constructed phylogenetic trees of the Trypanosomatidae family, and our data indicate that the molecular inferences do not sustain the traditional classification of these species. Our published findings are here reviewed along with recent unpublished observations of ribosomal RNA and 5S gene structures