EST sequencing from Triatoma infestans cDNA libraries.

AVILA ML, TEKIEL V, BUA J, SANCHEZ DO.

Carlos Paz, Cordoba, Argentina

Congreso; 44th Annual Meeting – Argentine Society for Biochemistry and Molecular Biology Research (SAIB).; 2008

Argentine Society for Biochemistry and Molecular Biology

EST sequencing from T. infestans cDNA librariesTriatoma infestans is one of the main vectors of Chagas disease in Argentina and South America. In order to contribute to the generacion of its transcriptome, we are constructing cDNA libraries to obtain Expressed Sequence Tags (ESTs) from IVth instar nymphs.In a first attempt, we constructed a standard cDNA library. The obtained sequences were compared against the non-redundant nucleotide collection of the NCBI database using the BLAST tool. So far, we have sequenced and analyzed 131 ESTs. We obtained 37% of sequences with non-significant similarity to any known sequence, and 12% with significant similarity to mRNAs of other organisms. These two subsets of ESTs are interesting for gene discovery and will be further analyzed. We also obtained 22% of sequences with significant similarity to rRNAs of T. infestans or other triatomine species, and 27% of sequences with identity with Triatoma virus, a RNA virus widely distributed in T. infestans populations. Clustering of these ESTs, revealed that most of them were redundant. Finally, 2% of the sequences had identity with mRNAs of T.infestans already annotated.With the aim of increasing the proportion of ESTs from low abundance transcripts, we are currently generating normalized cDNA libraries using the Clontech PCR-Select cDNA Subtraction kit. The comparative analysis of these libraries will be shown in the poster.