Our Project aims to engineer a new protein able to produce an immunological response and could be the base for a new type of vaccine based in Synthetic Biology techniques. We took advantage of Bioinformatics, used several databases featuring protein sequences and 3D structures in order to learn differences and similarities among the proteins of the 4 serotypes of the dengue virus, to recognize the antigenic determinants called epitopes and search for them in the correspondent databases.
We have designed an ideal protein containing epitopes from all 4 serotypes, and four more, each one containing just one of the 4 serotypes. We searched on protein databases for a suitable protein that bacteria normally exports from the cell, that has high levels of expression in our selected cell lineage, monomeric in order to take just one gene to be synthesized, big enough to contain several loops to hold all four epitopes and not too big or complicated so bacteria can synthetize it properly. We also took advantage of software and servers in order to check 3D properties of the final structure. Finally, we made our construct including promoter, RBS the main new sequence and other termination factors.
We ordered the synthesis of the respective coding genes, to make 5 biobricks, one containing the four serotypes and the other 4 just one. Later in the summer we used the biobricks to transform our bacteria strain CD41 and check the expression of the designed protein. After we accomplish expression and isolation of the proteins we will make bioessays for effectiveness, using sera from patients recovered from dengue, then with lab animals and finally with human volunteers.
