Fig. 1.

Construction of the two partial cDNA clones. A schematic drawing of the WNV genome is indicated on the top. Dark gray boxes represent the viral structural and light gray boxes the nonstructural part of the coding sequence. The 5′ and 3′ noncoding regions (NCR) are indicated as thin lines. (a) Six WNV sequence segments were generated by gene synthesis (vector backbones are not shown). The 5′TL AB segment is corresponding to WNV genomic sequence nt 1–1792, 5′TL CD to nt 1789–3632, segment 3′TL AB to nt 3622–5801, 3′TL CD to nt 5792–8028, 3′TL EF to nt 8022–10,025 and 3′TL GH to nt 10,022–11,029. (b) Fragments 5′TL AB and 5′TL CD were ligated yielding the final 5′ partial clone insert (5′TL) and two 3′ part fragments were generated by fusing fragments 3′TL AB with 3′TL CD and 3′TL EF with 3′TL GH. (c) Final partial clones, integrated in pBR322, are indicated. The 3′ partial clone was generated by ligation of the 3′TL ABCD with the 3′TL EFGH insert, taking advantage of the unique SpeI restriction site.