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. 2016 Jun 1;11(6):e0156579. doi: 10.1371/journal.pone.0156579

Fig 1. DTMUV reverse genetics strategy.

Fig 1

(A) Cloning design. Total RNA was extracted from the DTMUV FX2010-180P strain and reverse-transcribed into cDNA. Six cDNA segments were amplified using High Fidelity DNA polymerase pfx and inserted into plasmid vectors. The three 3’ cDNA segments were sub-cloned into plasmid p3656-10991. To generate the full-length DNA corresponding to the entire viral genome, a series of PCRs was performed using using pfu Ultra II Fusion HS DNA polymerase with the recombinant plasmids as templates. (B) Genome schematics of rFX2010-180P and rFX2010-180P-EGFP. rFX2010-180P-EGFP expresses eGFP from an IRES located between the NS5 gene and the 3’ UTR.