TABLE 1.
Properties of VP4 and NSP3 mutant virusesa
| Viruses |
Protein products of mutated gene |
||||||
|---|---|---|---|---|---|---|---|
| Name | Titers (PFU/mL) | Total size of VP4 gene (bp) | Rescue success | VP4 size (aa) | VP4 size (kDa) | VP4 cleaved products |
|
| VP5* size (kDa) | VP8* size (kDa) | ||||||
| SA11 WT | 1.65E + 08 | 2,331 | 3/3 | 776 | 86.1 | 60 | 28 |
| HR2 | 2.25E + 06 | 2,430 | 809 | 89.8 | 31 | ||
| NTD | 4.75E + 06 | 2,400 | 799 | 88.7 | 30 | ||
| RBM.1 | 2.75E + 06 | 2,400 | 799 | 88.7 | 30 | ||
| RBM.2 | 2.50E + 06 | 2,412 | 803 | 89.1 | 30 | ||
|
Name |
Titers (PFU/mL) |
Total size of NSP3 gene (bp) | Rescue success | NSP3 size (aa) | NSP3 size (kDa) | T2A product/s (aa) | T2A product/s (kDa) |
| RF WT | 1.30E + 08 | 942 | 3/3 | 313 | 34.7 | n/a | n/a |
| RBM | 1.10E + 08 | 1,158 | 2/3 | 385 | 42.7 | ||
| T2A_RBM | 9.20E + 07 | 1,212 | 3/3 | 403 | 44.7 | 330 + 73 | 36.6 + 8.1 |
| RBD | 1.88E + 07 | 1,521 | 1/3 | 506 | 56.1 | n/a | n/a |
| T2A_RBD | 7.70E + 07 | 1,575 | 3/3 | 524 | 58.1 | 330 + 194 | 36.6 + 21.5 |
a
The hypervariable region of SA11 VP4 was tagged with SARS-CoV-2 spike peptides: heptad repeat 2 (HR2), N-terminal domain (NTD), and receptor binding motif (RBM) regions. The C terminus of RF nonstructural protein (NSP) was tagged with either the RBM or receptor binding domain (RBD) of SARS-CoV-2 spike with or without a Thosea asigna virus 2A (T2A) peptide; WT, wild type; n/a, not applicable.