Heterologous expression in Pichia pastoris of gene BMTI optimized

The cattle tick, Rhipicephalus microplus, is the main tick species that undermines productivity losses in livestock throughout Brazil. Its control is primarily chemical and faces problems due to the resistance of ticks to several active principles used. The immune control is a viable alternative to tick control, and the protein BMTI is a strong candidate as a vaccine antigen. The BMTI sequence was cloned in eukaryotic expression vector pPICZ alpha A, respecting the make reading the AOX1 promoter. The resulting plasmid, pPICZ alpha BmTIS, was transformed into X33 strains of P. pastoris. A clone (X33BmTIS17) was sequenced and selected to be induced. SDS-PAGE showed a major band with the approximate size of 49 kDa and in Western bloting was recognized by the anti-histidine and by bovine serum immunized with a synthetic peptide (BmTI).