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. 1998 Mar;72(3):2364–2372. doi: 10.1128/jvi.72.3.2364-2372.1998

FIG. 5.

FIG. 5

The 5′-coding region of Npro is essential for autonomous RNA replication of DI9c. (A) Top: schematic drawing of the DI9c derivatives used in this study (see the legend to Fig. 1A for details). As indicated (MG1590) and as described in detail in Materials and Methods, DI9cNS3 RNA contains an additional AUG codon upstream of GGA coding for glycine 1590 of BVDV CP7 NS3. DI9cubi RNA also contains the ubiquitin gene (ubi), and DI9cΔNproubi also contains the initial 42 codons of the Npro gene (indicated by Δ). Bottom: RNA and primary amino acid sequence comparisons for the initial five codons of BVDV CP7 cRNA and the DI9cubi ORF. (B) RNase protection assay of BHK-21 cells 24 h p.t. with different DI9c derivatives. Lanes: 1, protection of RNA derived from cells transfected with DI9c (positive control); 2, protection of RNA p.t. with DI9cNS3; 3, protection of RNA p.t. with DI9cubi; 4, protection of RNA p.t. with DI9cΔNproubi; p, sense probe (1/10 of the experimental input was loaded on the gel). (−), sense probe; (+), antisense probe.