Fig. 2

Cloning strategy to incorporate the eGFP gene into the full-length NDV clone rLS1. Three fragments ligation reaction was performed to insert the eGFP gene into the rLS1 genome between AsiSI and BbvCI restriction sites. Fragment-A containing the eGFP cassette was chemically synthetized, whereas, fragment-B was amplified by PCR using the pFLC-LS1 clone as a template. Both fragments were simultaneously cloned into the pFLC-LS1 to generate the pFLC-LS1-1eGFP. The final construct includes the eGFP ORF, flanked by gene-start and gene-end from the N gene, inserted at the 3′- proximal position of the NDV genome