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. 2018 Apr 24;10(5):216. doi: 10.3390/v10050216

Figure 3.

Figure 3

Viral infection and GFP expression of LIYV-based “ORF substitution” expression vector in Hc-Pro Nicotiana benthamiana plants. (A) LIYV symptoms in upper non-inoculated leaves of LIYV-L2-P5ΔGFP infected plants at 3 wpi and GFP fluorescence visualized under UV light. (B) Detection of viral infection and insertion integrity by RT-PCR using total RNA extracted from upper non-inoculated leaves of LIYV-WT and LIYV-L2-P5ΔGFP agroinoculated plants. Two primer sets were used to amplify the sequence of LIYV CP (CP, 530 bp) and the sequence flanking the GFP ORF (Insert, 882 bp), LIYV-WT without the GFP ORF was used as a control (282 bp). (C) Immunoblot analysis of LIYV CP and GFP accumulation in upper non-inoculated leaves infected by LIYV-WT, LIYV-L2CGFP, and LIYV-L2-P5ΔGFP using LIYV CP and GFP specific antibodies. The Ponceau S stained rubisco large subunit serves as a loading control.