TABLE 1.
Details of PB1-F2 mutants made in this studya
| Source virus | Name of mutant | PB1-F2 status or size (aa) | Mutagenesis primer(s) |
|---|---|---|---|
| VN/281 | VN/281/PB1-F2FL | Full length (90) | Kpn1PB1F76-94 and Xba1PB1R384-67 |
| VN/281/PB1-F2TR | Truncatedb (24) | PB1C167T_C170T_F | |
| VN/281/PB1-F2NO | Deletedc | PB1T96C _F | |
| VN/281/PB1-F2FL-HA | Full length (90) | Kpn1PB1F68-90 and Xba1HAtagPB1R364-43 | |
| VN/281/PB1-F2HA-TR | Truncatedb (24) | PB1F151-185 (HA_C167T_C170T) | |
| VN/281/PB1-F2TR-HA | Truncatedd | PB1C167T_C170T_F and Xba1HAtagPB1R364-43 | |
| VN/281/PB1-F2NO-HA | Deletedc | PB1T96C _F | |
| VN/296 | VN/296/PB1-F2FL | Full length (90) | PB1C96T_F |
| VN/296/PB1-F2TR | Truncated (24) | PB1C167T_C170T_F | |
| VN/296/PB1-F2NO | Deletedc | Kpn1PB1F76-94 and Xba1PB1R384-67 | |
| VN/296/PB1-F2FL-HA | Full length (90) | PB1C96T_F | |
| VN/296/PB1-F2HA-TR | Truncatedb (24) | PB1F151-185 (HA_C167T_C170T) | |
| VN/296/PB1-F2TR-HA | Truncatedd | PB1C167T_C170T_F and Xba1HAtagPB1R364-43 | |
| VN/296/PB1-F2NO-HA | Deletedc | Kpn1PB1F68-90 and Xba1HAtagPB1R364-43 |
a
A total of 6 mutants were made from each virus: 3 with a C-terminal HA tag and 3 without the HA tag. See Fig. 1 for schematic diagrams.
b
Stop codons were added at positions 25 and 26; the plasmid is expected to express a truncated protein (24 aa). One-half of these mutants have the HA tag immediately upstream from the stop codons.
c
In this variant, only the first ATG is mutated to ACG, leaving the downstream in-frame ATGs intact.
d
This variant has stops at positions 25 and 26, but the HA tag is at the C terminus.