FIG 1.

Schematic overview of chimera insert sizes and locations and sequence alignment of donor EIII sequences. (A) Donor EIII sequences were cloned into the WNV infectious clone using four combinations of up- and downstream restriction sites. The four resulting constructs were labeled A, B, C, and D. The A forms had the largest insertion size, with the donor virus EIII sequence being flanked by the donor virus EI sequence upstream and the first stem-helix structure downstream. The B forms contained donor virus EIII without the additional sequences, while the C and D forms contained the extra up- and downstream sequences, respectively. EH, E helical domain; TM, transmembrane domain. (B) Donor virus EIII amino acid sequences were aligned to determine the differences present in the chimeric viruses. Amino acid position numbers correspond to those of the WNV NY99 E sequence. Alignments were performed using the Geneious (v9.0.2) program (Biomatters, Auckland, New Zealand).