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 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.
Toma
Helena K
HK
Departamento de Análises Clínicas e Toxicológicas, Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, Av. Brigadeiro Trompowsky s/n, 21941-590 Rio de Janeiro, RJ, Brazil.
Yamada-Ogatta
Sueli F
SF
Brandão
Adeilton
A
Krieger
Marco A
MA
Goldenberg
Samuel
S
Fernandes
Octavio
O
D000818:Animals; D001483:Base Sequence; D044822:Biodiversity; D015139:Blotting, Southern; D001938:Brazil; D003001:Cloning, Molecular; D016000:Cluster Analysis; D016054:DNA, Protozoan / Q000032:analysis / Q000737:chemistry; D048293:Didelphis; D004587:Electrophoresis, Agar Gel; D005819:Genetic Markers; D014644:Genetic Variation; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012239:Rhodnius; D014349:Trypanosoma cruzi / Q000145:classification* / Q000235:genetics
D016054:DNA, Protozoan; D005819:Genetic Markers