Figure 5. Recombination of complementing viruses after serial passaging.

(A) PCR strategy to detect gene arrays with outward-facing primers. (B) Representative PCR products amplified with outward-facing primers for VC-R2 and three independent HGT3 passages. (C) Recombination of complementing viruses resulted in tandem arrays of the H4L locus as determined by long-read sequencing.

Figure 5—figure supplement 1. Recombination sites in passaged HGT3.

(A) Recombination site in HGT3 between H7R and H3L identified in passage 10 of replicate 3 and passage 17 of replicate 2 by PCR and Sanger sequencing, as well as in passage 17 of replicate 3 by PacBio sequencing. The red rectangle shows three shared nucleotides between the genes at the breakpoint. (B) Recombination site in HGT3 between H6R and H3L identified in passage 2 of replicate 3 by PCR and Sanger sequencing. The red rectangle shows three shared nucleotides between the genes at the breakpoint. (C) Schematic representation of recombined loci in parental VC-R2 and HGT3, and recombined loci in r3P17 (blue dotted line) and r3P2 (red dashed line).