Figure 5.

In vitro Bamboo mosaic virus (BaMV) RNA template activity assay of CMV/263‐derived mutants. (A) A diagram of the various constructs derived from the entire 3′ untransformed region of CMV RNA (CMV/263). (B) mFOLD‐predicted secondary structure of the 83‐nucleotide fragment of CMV/263 fused with an extra UUUUC sequence at the 3′‐end. (C) In vitro RNA‐dependent RNA polymerase (RdRp) template activity assay. For RNA transcripts of Ba‐77 and the CMV/263‐derived constructs, products of the BaMV RNA template activity assay were labelled with [α‐³²P]UTP, separated by polyacrylamide gel electrophoresis and detected by autoradiography. Quantification of the RdRp template activity assay products. The intensities of the bands detected in (B) were quantified. The level of the Ba‐77 product was arbitrarily designated as 100 and used to normalize the other signals. Data presented are the average and error bars of at least three independent reactions.