Cloning of the chimera gene tv70catl of trypanosoma vivax in a bacterial system

Abstract

Animal trypanosomosis is a hemotropic parasitic disease caused mainly by Trypanosoma vivax (T. vivax), which causes significant economic losses in Venezuelan livestock. To date, the serodiagnosis of the disease is limited by the absence of specific antigens for the identification of T. vivax. The cloning of T. vivax chimera genes has not been reported so far and could be the key in the development of immunoprotective strategies for the control of this disease. A chimera gene constituted by the binding of the catalytic region of the Cistein protease to the region of the Hsp70359-610 of T vivax was cloned. For this, the pPiczαA vector was extracted and digested with restriction enzymes EcoRI and NoTI, obtaining the release of the Tv70CatL gene of 1478 bp. Subsequently, it was ligated to the expression vector pET28a and transformed into strains of E. coli Bl21 key in the development of immunoprotective strategies for the control of this disease. A chimera gene constituted by the binding of the catalytic region of the Cistein protease to the region of the Hsp70359-610 of T vivax was clo
ned. For this, the pPiczαA vector was extracted and digested with restriction enzymes EcoRI and NoTI, obtaining the release of the Tv70CatL gene of 1478 bp. Subsequently, it was ligated to the expression vector pET28a and (DE3) by electroporation. The colonies were analyzed by PCR, with amplification being positive. Then, the product was digested with restriction enzymes EcoRV and PstI thus verifying the sequence of the recombinant molecule.