Molecular Studies of Glycerol-3-phosphate dehydrogenasefrom Triatoma infestans

STROPPA M, SORIA N., CARRIAZO C., PEREIRA R. Y GEREZ DE BURGOS N.

Mar del Plata (Argentina)

Congreso; XLIII Reunión Anual de SAIB; 2007

Sociedad Argentina de investigación en bioquímica y Biología Molecular

Triatoma infestans (T. infestans), Chagas' disease vector, acquires wings and the ability to fly after the last molt from fifth instar nymph to adult. The ability to fly is important for insect dispersion. In previous studies, in our laboratory, it was demostrated that glycerol-3-phosphate dehydrogenase (GPDH), which is involved at glycerophosphate shuttle, increase its activity 30 fold in adults thoracic muscles. Aduls muscles should have higher glycolitic and respiratory capacity to support fly activity. Electrophoretic studies from thoracic muscles showed two GPDH isoenzymes. Nymphs predominant isoform has less mobility. The aim of this work is to begin GPDH molecular studies. Using cDNA pools with degenerated primers, we amplified a 200 bp RT-PCR product from 1 to 3 days old T. infestans adults thoracic muscles. Upong sequencing and database alignments the fragment shows to be part of GPDH cDNA. Based on the sequence obtained, specific primers were designed for RACE experiment. WE successfully amplified completes cDNA of two GPDH isoforms. Sequencing results allow identity, homology and conserved domains comparative studies with others species. GPDH isoform expression, studied by RT-PCR showed stage and tissue specific pattern.