Figure 1
Design of infectious cDNA clones of DENV3 and generation of recombinant DENV4/3 viruses. (A) Genome schematic of DENV4 infectious clone design including restriction endonucleases used to generate subgenomic fragments from individual high-copy plasmids. Size of subgenomic fragments indicates positions in DENV genome where breaks were made to circumvent bacterial instability and toxicity. (B) Amino acid residues of a single 5J7 epitope are shown teal balls. In each ribbon structure of E protein, Domain 1 is red, Domain 2 is yellow and Domain 3 is blue. Residues of the 5J7 epitope spread across all three monomers (B, B’ form a dimer, A is in the adjoining monomer) are teal. PDB accession code 4CBF(DENV4). (C) Four E monomers in a partial raft are shown for each 5J7 transplant with increasingly larger transplants into DENV4. Transplanted amino acids in each of the DENV4 chimeras can be seen as highlighted AA; DENV4 M12 (green residues), DENV4 M14 (green + cyan residues), and DENV4 M16 (green + cyan + orange residues). (D) The chart shows the amino acids changed in DENV4 E protein by transplantation of DENV3 amino acid sequences to generate DENV4 M12, M14, and M16 respectively. Amino acid number represents residue from start of DENV4 E protein. The K323Q mutation in DENV4 M16 is a tissue culture adaptation and is included in the orange balls in B. Colors correspond to highlighted AA residues in (C).
