Construção de candidatos a vacinas de DNA baseados nos genes prM e envelope do Zika vírus

Directly associated with the outbreak of fetal malformations and the increase of cases of adults affected with Guillain-Barre Syndrome in Latin America, and especially in Brazil, Zika virus (ZIKV) is the subject of several researches worldwide. Like Dengue and Yellow Fever viruses, ZIKV belongs to the Flaviviridae family and has as main transmission vector the mosquitoes from the genus Aedes. But also, it can be transmitted by other routes such as sexual relations and body fluids. Thus, in view of its rapid spread, the development of prophylactic and therapeutic strategies against ZIKV is extremely important. Currently, there are no prophylactic vaccines available against ZIKV. In this sense, in this work we developed candidates for DNA vaccines based on the prM and Envelope genes of ZIKV, which were selected for the humoral immune response that they can elicit. Three constructs were developed in this study: the first containing the complete Envelope (E) sequence, the second containing only the last 34 amino acids of the C-terminal region of prM and the complete Envelope sequence (prM34.E), and the third was Envelope with a deletion of the last 108 amino acids of the C-terminal region (EnvΔ108). All constructs were cloned into the pGEM-T easy vector, and then subcloned into the expression vector for mammalian cells pVAX1 and transfected into Vero cell cultures. After the transfections, detection of proteins of interest was performed by western blot and the verification of gene transcription was done by RT-PCR. The results obtained demonstrated expression of E, although expression of prM34.E and EnvΔ108 was detected. Similarly, only E protein detection was possible, and protocol optimizations are required for future detection. The efficacy of the E-construct as a vaccine candidate should be evaluated in future animal immunological assays.