Figure 2.

Schematic representations of the MVs rescued from the plasmid molecular clones used to derive MV-NIS: p(+)MV-tag produced MVtag; p(+)MHlrGFPV produced MHlrGFPV; and p(+)MH-NIS(HP)3 produced the drug substance MV-NIS. To construct p(+)MH-NIS(HP)3, the MluI–AatII fragment of p(+)MHlrGFPV containing the additional gene encoding rGFP was replaced with a PCR product containing the human NIS gene flanked by MluI and AatII sites.¹² An artificial gene boundary region (nucleotide sequence shown) was inserted into the SpeI site (position 9234) using two synthesized oligonucleotides. The p(+)MHlrGFPV plasmid was constructed by inserting the rGFP coding region into the MluI and NruI cloning sites of the polylinker. The AatII site is in the rGFP 3′-non-coding sequence. The p(+)MH-NIS(HP)3 plasmid was constructed by inserting the human NIS coding region into the MluI and AatII sites of p(+)MHlrGFPV, replacing the rGFP gene. The position of the gene coding regions is indicated above the MVtag genome. To facilitate the comparison of the MV-NIS genome sequence (17,940 nucleotides) with other MV genomes of the Edmonston lineage (15,894 nucleotides), the 2,046 bp inserted transcription unit containing the NIS coding region is numbered separately (1N–2046N).