Cloning and characterization of the gene encoding the heat shock protein HSP83 from Trypanosoma cruzi

[Objective] Tryapanosoma cruzi, causal agent of Chagas' disease, is a parasite that presents morphological alternation between an invertebrate (triatomine) and a vertebrate (mammal) host. Several studies have shown that in T. cruzi, HSP83 (homolog of HSP90) is essential for cell division and control of the response to thermal stress. The objetive of the present work was the cloning, bioinformatics characterization and expression of the heat shock protein HSP83 T. cruzi gene. [Methodology] RNA extraction was performed from epimastigotes using a commercial kit. The cDNA encoding HSP83 was obtained by RT-PCR, from the extracted mRNA, for which the primers were designed based on the HSP83 sequence of T. cruzi strain CL Brener. Cloning was performed on pGEM®T-Easy, and subcloned into the expression vector pQE30. Sequence and bioinformatics characterization was performed. The gene was expressed and the recombinant protein was purified by affinity chromatography and identify by immunoblotting. [Results] Sequence analysis showed similarity to the gene encoding HSP83 from Trypanosoma cruzi and HSP domains were observed, as well as B epitopes in the sequence. After 3 hours of induction with IPTG a recombinant protein with an approximate weight of 83 kDa was obtained. The immunoblotting reaction with hyperimmune anti-T. cruzi epimastigote serum allowed the detection a single band with a molecular weight of approximately 83 kDa [Conclusions]. All results indicate that the cloning and characterization of HSP83 from Trypanosoma cruzi was achieved