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. 2024 Mar 7;16(3):411. doi: 10.3390/v16030411

Figure 2.

Figure 2

Production of recombinant OSU that encodes a foreign protein. (A) Modifications of rotavirus genome segment 7. The schematics indicate the nucleotide positions of the coding sequences for NSP3, the porcine teschovirus 2A element, 3X FLAG, and the fluorescent reporter UnaG (green). The red arrows indicate the positions of the 2A translational stop-restart elements, and the asterisks indicate the ends of the open reading frames. (B) Recovery of recombinant OSU-2A-UnaG via reverse genetics. Viral dsRNAs from OSU-tc_(MA104), rOSU, rOSU-2A, and rOSU-2A-UnaG were resolved via electrophoresis on an 8% polyacrylamide gel and stained with ethidium bromide. The migrations of modified genome segment 7 are indicated with red arrows. Genome segments 1–11 of rOSU are indicated on the left side of the panel. (C) Genetic stability. rOSU-2A and rOSU-2A-UnaG were serially passaged on MA104 cells. Viral dsRNAs from a total of five passages (P) were resolved via electrophoresis on an 8% polyacrylamide gel and stained with ethidium bromide. The migrations of modified genome segment 7 are indicated with a red arrow. Genome segments 1–11 of rOSU are indicated on the left side of the panels.