In Silico Vaccine Design and Expression of the Multi-Component Protein Candidate against the Toxoplasma gondii Parasite from MIC13, GRA1, and SAG1 Antigens

Background:We aimed to design a B and T cell recombinant protein vaccine of Toxoplasmagondii with in silico approach. MIC13 plays an important role in spreading the parasite in thehost body. GRA1 causes the persistence of the parasite in the parasitophorous vacuole.SAG1 plays a role in host-cell adhesion and cell invasion.Methods:Amino acid positions 73-272 from MIC13, 71-190 from GRA1, and 101-300from SAG1 were selected and joined with linker A(EAAAK)A. The structures, antigenicity,allergenicity, physicochemical properties, as well as codon optimization and mRNA structureof this recombinant protein called MGS1, were predicted using bioinformatics servers. Thedesigned structure was synthesized and then cloned in pET28a (+) plasmid and transformedinto Escherichia coli BL21.Results:The number of amino acids in this antigen was 555, and its antigenicity was esti-mated to be 0.6340. SDS-PAGE and Western blotting confirmed gene expression and suc-cessful production of the protein with a molecular weight of 59.56kDa. This protein will beused in our future studies as an anti-Toxoplasma vaccine candidate in animal modelsConclusion:In silico methods are efficient for understanding information about proteins,selecting immunogenic epitopes, and finally producing recombinant proteins, as well as re-ducing the time and cost of vaccine design.