Fig. 2

Plaque of PRV B-mini-F, PRV B-gD^S-mini-F and PRV B-gD&gC^S, RFLP of pB-mini-F, pB-gD^S-KAN-mini-F and pB-gD^S-mini-F, and PCR verification of gC and gD genes replacement. A Images of PRV B-mini-F, PRV B-gD^S-mini-F and PRV B-gD&gC^S plaques under UV excitation and contrast are shown. B DNA from pB-mini-F BAC clone (lanes 1 and 4) and recombinant BACs of pB-gD^S-KAN-mini-F (lanes 2 and 5) and pB-gD^S-mini-F (lanes 3 and 6) were prepared by mini-prep and digested with Hind III (lanes 1–3) or Sph I (lanes 4–6). Digests were separated by 0.8% agarose gel electrophoresis for 15 h under 40 V. Predictions of these digestions were performed using whole genome sequences of Bartha-K61 as a reference (GenBank ID: JF797217.1). C Verification of gC and gD genes replacement by PCR. gD of Bartha-K61 and PRV B-gD&gC^S were identified with AH02LA-gD-F/AH02LA-gD-R. gC of Bartha-K61 and PRV B-gD&gC^S were identified with SEQ-AH02LA gC F/SEQ-AH02LA gC R