TABLE 2.
Substitutions introduced into the HEF protein of C/JHB/1/66 and enzyme restriction strategy used to analyze the stability of the rescued mutants
| Virus description | Amino acid position | Amino acid | Nucleotide sequenceb | Restriction site | After digestionc |
|---|---|---|---|---|---|
| wta | 284 | Thr | GAC ACC | None | 875 |
| 465 | Thr | ACA GGA | None | 875 | |
| Mutants | 284 | Ile | GAT ATC | EcoRV | 694 + 181 |
| 465 | Ala | GCC GGC | NgoMIV | 727 + 148 |
a
Wild type.
b
Underlined, nucleotide codons coding for the indicated amino acid; in bold, nonsilent mutations; in italics, silent mutations introduced in the neighboring codons to design a restriction enzyme site.
c
Expected length(s) (nt) of fragment(s) produced after digestion of amplicons with the indicated restriction enzymes.