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Insertion of ORF2 in different orientations exerted distinct inhibitory effects on gene expression. Total RNA extracted from HeLa cells transfected with plasmids was analyzed by northern blotting. ORF2 in sense orientation inhibited GFP expression much more strongly than in antisense orientation; in the latter orientation ORF2 also caused premature transcriptional termination. Arrow on the left shows the expected positions of GFPORF2 and GFPlacZ that are of the same length.
