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. 2007 May 30;81(16):8396–8405. doi: 10.1128/JVI.00321-07

FIG. 2.

FIG. 2.

Adaptation of mutants 4 and 5 (Mut 4 and Mut 5), RICO-ΔZ, RICO-PV, CBV-RICO, and CBV-ΔZ through serial passage in Sk-N-Mc cells. (A) One-step growth curves of prepassage viruses (open bars) and viruses subjected to eight serial passages (gray bars). Individual constructs are indicated for the 0-h time point and are shown in the same order for the 6- and 8-h time points; CBV-RICO (RICO) is indicated in each set of bars. (B) Nucleotide (Nt) substitutions identified in the indicated viruses through sequencing of bulk PCR product and their effect on the amino acid sequence of the affected viral protein. Nucleotide numbering refers to the genome of CBV-RICO. Amino acid (aa) numbering refers to the residue within CBV3 3A or 3C. Adaptation mutations are shown as numbers and letters in brackets. (C) Chromatograms depicting sequencing results for mutant 4. Double peaks indicate the presence of adaptation mutations (the wt sequence is shown above the chromatograms); nucleotide numbering corresponds to that in panel B.