Table 1.
AmiB variants isolated in the plasmid release enrichment.
| Variants from Enrichmenta |
Lethal Activityf | |
|---|---|---|
| WT | ΔenvC | |
| W8L, N263D, M295Ib | +++ | +++ |
| G33D, E300Kb | +++ | +++ |
| S87P, S306L | +++ | +++ |
| G170C, S293Rb | +++ | +++ |
| S173N, S293R | +++ | +++ |
| L229S, A296V, D314Vb | +++ | +++ |
| S293Ic, e | +++ | +++ |
| S293Re | +++ | +++ |
| M295Ic, e | +++ | +++ |
| A296Vc, e | +++ | +++ |
| E300Kc, e | +++ | +++ |
| H302P, A405Vb, e | +++ | +++ |
| E303Ke | +++ | +++ |
| S306Lc, e | +++ | +++ |
| S306Pd, e | +++ | +++ |
| Q333Le | + | +++ |
| R372Ce | + | +++ |
| Site-Directed Variants | ||
| H302P | +++ | +++ |
| S306P | +++ | +++ |
| D314V | − | − |
| A405V | − | − |
a
each row in the table indicates the substitution(s) found in a single variant
b
also contain silent mutation(s)
c
variant isolated from two independent libraries
d
variant contains a premature stop at codon 422
e
lytic phenotype phenotype was reproduced upon subcloning into the parental plasmid backbone. All others were not subcloned.
f
Plasmids encoding the indicated AmiB variants under control of the arabinose promoter were assessed for lethal activity by streaking transformants of TB28 [WT] or TB140 [ΔenvC] on M9-CAA-Cm10 agar containing either 0.2% glucose or 0.2% arabinose and visually determining if growth was significantly inhibited on the arabinose-containing plate.