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: New York; 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 paper 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 ± 0.16mM and an estimated Vmax value of 476.2 nmoles/h/mg of protein. These 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 in a powerful tool?easy to follow by its fluorescence?to study the role of polyamines in Chagas disease pathology and in related processes such as parasite survival, invasion, proliferation,metacyclogenesis, and tissue spreading.