Trypanosoma cruzi: Subtractive hybridization as a molecular strategy to generate new targets to distinguish groups and hybrids

RAPD analysis and sequences of the mini-exon and ribosomal genes show that Trypanosoma cruzi can be clustered into two phylogenetic groups— T. cruzi I and II. Herein, the Representational Difference Analysis (RDA) method was used, providing new targets specific for each group. After three rounds of...

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Bibliographic Details
Published in:Experimental parasitology 2007-10, Vol.117 (2), p.178-187
Main Authors: Toma, Helena K., Yamada-Ogatta, Sueli F., Brandão, Adeilton, Krieger, Marco A., Goldenberg, Samuel, Fernandes, Octavio
Format: Article
Language:English
Subjects:
10 mM Tris–HCl pH 8.0, 10 mM EDTA, pH 8.0
Animals
Base Sequence
Biodiversity
Biological and medical sciences
Blotting, Southern
Brazil
Cloning, Molecular
Cluster Analysis
deoxyribonucleic acid
Didelphis
DNA
DNA, Protozoan - analysis
DNA, Protozoan - chemistry
Electrophoresis, Agar Gel
Fundamental and applied biological sciences. Psychology
Genetic Markers
Genetic Variation
Humans
Hybrids
Life cycle. Host-agent relationship. Pathogenesis
LIT
liver infusion tryptose
Molecular markers
Molecular Sequence Data
Nucleic Acid Hybridization - methods
PBS
phosphate buffer saline
Phylogenetic lineages
Phylogeny
Polymerase Chain Reaction
Protozoa
RDA
representational difference analysis
Rhodnius
Saline sodium citrate
SDS
sodium dodecyl sulphate
SSC
Trypanosoma cruzi
Trypanosoma cruzi - classification
Trypanosoma cruzi - genetics
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Summary:RAPD analysis and sequences of the mini-exon and ribosomal genes show that Trypanosoma cruzi can be clustered into two phylogenetic groups— T. cruzi I and II. Herein, the Representational Difference Analysis (RDA) method was used, providing new targets specific for each group. After three rounds of RDA hybridizing F strain (tester) with Y strain (driver) and vice-versa, an F-specific (F#30) and Y-specific (Y#22) clone were obtained specifically recognizing isolates from Amazonas ( T. cruzi I) and Piauí ( T. cruzi II). These segments corresponded to an unspecified protein (F#30) and a trans-sialidase (Y#22). Analysis of the F#30 sequence in T. cruzi I, T. cruzi II and zymodeme 3 samples displayed negligible specific differences that distinguished each group. In addition this F#30 gene has great potential as a hybrid marker.
ISSN:0014-4894
1090-2449
DOI:10.1016/j.exppara.2007.04.008