Single and multiple nucleotide deletions within the 17 nt functional region and detection of viral RNAs in protoplasts.

<p>(A) The 17 nt functional region (nt 2601–2617) of wtTNV-A^C (pMTC 27) is shown on top, and the arrow indicates the translation direction from the +1 start site of the CP gene to the +6 position. Nucleotides deleted in the mutants are indicated by dashed lines. (B) Viral RNA accumulation in tobacco BY-2 protoplasts transfected by the mutants was assessed by Northern blots at 18 hpt with the same probe used in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057938#pone-0057938-g001" target="_blank">Fig. 1</a> and the CP (P30) was identified in Western blots with a antibody raised against the TNV-A^C CP. The TNV-A^C RNA species and CP (P30) are indicated on the left side of the gel photographs and plant rRNAs are shown as loading controls in the bottom panels.