Figure 9. The priming activity of ApG.

A. The effect of biotin-labeled ApG (Bio-ApG) on BTV polymerase was compared with non-labeled ApG (ApG). Each 20 pmol of ApG or Bio-ApG was added to the reaction mixture containing [α-³²P] CTP. B. Priming activity by dinucleotide, ApG, was detected using biotin-labeled ApG and non-radioactive polymerase assay as described in Materials and Methods. As markers, BTV genomic segments was used and stained with 0.02% (w/v) methylene blue after transferring to the nylon membrane (lane 1). Non-labeled ApG (lane 2) or biotin-labeled ApG (lane 3) was added the reaction mixture containing non-labeled rNTP with uncapped T7 S9 ssRNA. Biotin-labeled dsRNAs were detected using Streptavidine-alkaline phosphatase conjugate as described in Materials and Methods. Genome segments are indicated on the left.