Fig. 4

Generation of recMVA-NoV using marker-based approach. a After transfection of the pIIIH5-GII4VP1 into MVA-infected DF-1 cells, mCherry-expressing cells producing recMVAs were picked and plaque purification was carried out. After some rounds of plague picking, a significant increase in the signal was observed. Scale: 100 μm. b Confirmation of gene insertion into the MVA genome by PCR. After extraction of viral DNA from infected DF-1 cells, insertion of the NoV capsid gene was checked by amplifying the DelIII cassette with gene-specific primers. An amplified fragment of 1.6 Kb was expected for the GII.4. DF-1 cells infected with WT-MVA were used as negative control for PCR. c Confirmation of WT-MVA clearance. The presence of the WT-MVA was checked by amplifying DelIII cassette where a PCR product of 446 bp was expected