Trypanosoma cruzi co-expressing ornithine decarboxylase and green fluorescence proteins as a tool to study the role of polyamines in Chagas? disease pathology

BARCLAY JJ; MOROSI LG; VANRELL MC; TREJO EC; ROMANO PS; CARRILLO C

Enzyme Research

SAGE-Hindawi

Lugar: Londres; Año: 2011 p. 657460 - 657470

2090-0414

Polyamines are essential for Trypanosoma cruzi, the causative agent of Chagas´ disease. As T. cruzi behaves as a natural auxotrophic organism it relies on host polyamines biosynthesis. In this work we obtained a double-transfected T. cruzi parasite that expresses the green fluorescent protein (GFP) and a heterologous ornithine decarboxylase (ODC), used itself as a novel selectable marker. These autotrophic and fluorescent parasites were characterized; the ODC presented an apparent Km for ornithine of 0.51 ± 0.16 mM and an estimated Vmax value of 476,2 nmoles/h/mg of protein. This expressing ODC parasites showed higher metacyclogenesis capacity than the auxotrophic counterpart, supporting the idea that polyamines are engaged in this process. This double-transfected T. cruzi parasite results a powerful tool to study the role of polyamines, easily follow by its fluorescence, in Chagas? disease pathology and in related processes such as parasite survival, invasion, proliferation, metacyclogenesis and tissue spreading.