Skip to main content
. 2010 Jun;16(6):1138–1145. doi: 10.1261/rna.1962010

FIGURE 2.

FIGURE 2.

(A) Schematic representation of the BUNV S segment genome and its RNA synthesis activity, which is both transcription and replication. The annealing position of oligonucleotide MRP used to detect the 5′ end of these RNAs is shown. (B) Schematic representation of model BUNV segment G-DBL(0/0). This template was derived from BUN-S(ren) by the insertion of an additional genomic 3′ NTR, resulting in two identical 3′ NTRs, where RNA synthesis could initiate, designated NTRi (internal) and NTRe (external). (C) Identification of the position where replication and transcription initiated on the genomic strand of G-DBL(0/0). Model segment G-DBL(0/0) was expressed in BHK-21 cells along with BUNV S and L ORFs (+), or the S ORF alone (−). The resulting BUNV RdRp-specific RNAs were analyzed by primer extension using end-labeled oligonucleotide MRP. Labeled DNA fragments were subjected to PAGE on a sequencing gel, and bands corresponding to BUNV RNAs are marked with arrowheads. To act as size markers, RNAs generated by parental template BUN-S(ren) were analyzed alongside, and the corresponding cDNA was sequenced using MRP.