Figure 3
From: Engineering a vector-based pan-Leishmania vaccine for humans: proof of principle
Engineering a multi-sand fly species based anti-Leishmania vaccine. A single DNA-based vaccine was engineered, considering a reverse vaccinology approach applied to the two salivary proteins this study focuses on: PdSP15 and LJL-143. The scheme of the final DNA vaccine, consisting of the VR2001-TOPO vector backbone and the fusion salivary antigen coding sequence, composed by the selected immunogenic portions of the two salivary proteins joined contiguously in an interpolated manner, a FLAG-tag, a HIS-tag and a stop codon is represented (A). The final sequence coding for the fusion salivary antigen (codon optimized for the mammalian codon usage bias), used in the cloning approach, as well as the respective translation, is shown (B). The underlined nucleotides code for the portions of PdSP15, while the non-underlined ones code for the portions of LJL-143. FLAG- and HIS-tags coding sequences are highlighted. The fusion protein amino-acid sequence was subjected to in silico predictions of antigenicity, solubility upon overexpression, presence of transmembrane domains, disulphide bonds formation and allergenic potential, whose quantitative and/or qualitative results are represented (C).
