Gene Discovery Stage-Specific and Gene Expression Profiling in Trypanosoma cruz

MINNING, T; MCGRAW, RA; DOWLESS, D; GARG, N; BUA, J; GARCIA, GA; SHIFFLET, A; SANCHEZ, DO; TARLETON, RL

Athens, Georgia, Estados Unidos

Simposio; The University System of Georgia Research Symposium, Genomics: Research and Application; 2000

University of Georgia

There are no vaccines for Chagas' disease and specific chemotherapy for T. cruzi, the causative agent this human disease, is far from ideal.  Since only a small fraction of the T. cruzi genome has been sequenced, gene discovery efforts for the identification of vaccine candidates and chemotherapeutic targets would be facilitated by HTS approaches.  This ongoing study exploits recent advances in DNA microarray technology as a gene discovery platform.  DNA microarrays of 4,400 targets were constructed from T. cruzi ORF-selected and random, genomic sequencing libraries.  These microarrays were probed with mixtures of labeled cDNAs prepared from T. cruzi trypomastigotes (extracellular, non-dividing forms) and amastigotes (intracellular, dividing forms) to enable simultaneous, pairwise comparisons of the relative expression levels for the genes represented in the arrays.  It is hypothesized that the products of parasite genes expressed soon after host cell invasion may facilitate the recognition and destruction of infected cells by host CTLs.  Clones representing genes upregulated early in trypomastigote to amastigote differentiation will be selected for sequencing and genetic immunization studies.