Table 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant descriptiona | Source or reference |
|---|---|---|
| Escherichia coli strains | ||
| DH5α | F− supE44 hsdR17 recA1 gyrA96 endA1 thi-1 relA1 deoR λ− | 22 |
| JW3805 | F− Δ(araD-araB)567 ΔlacZ4787(::rrnB-3) λ− rph-1 ΔmetE774::kan Δ(rhaD-rhaB)568 hsdR514 | Keio Collection (National Institute of Genetics, Japan) |
| WE | JW3973 carrying the metAW3110 gene | 24 |
| WEΔmetE | WE ΔmetE::kan | This study |
| WE-214 | WE carrying mutated metE-214 gene | This study |
| WE-CA | WE carrying metE gene with C645A substitution | This study |
| WE-214A | WE carrying metE-214 gene with C645A substitution | This study |
| A39 | WE carrying metE gene with V39A substitution | This study |
| C46 | WE carrying metE gene with R46C substitution | This study |
| I106 | WE carrying metE gene with T106I substitution | This study |
| E713 | WE carrying metE gene with K713E substitution | This study |
| BL21(DE3) | F− ompT hsdSB(rB− mB−) gal dcm (DE3) | Novagen (Billerica, MA) |
| Plasmids | ||
| pKD46 | λ Red (gam bet exo) araC rep101(Ts), Apr | 23 |
| pET22b/MetE | Expression vector contains wild-type metE gene, Apr | This study |
| pET22b/MetE214 | Expression vector contains mutated metE-214 gene, Apr | This study |
| pET22b/MetCA | Expression vector contains metE gene with C645A substitution, Apr | This study |
| pET22b/MetE214A | Expression vector contains mutated metE-214 gene with C645A substitution, Apr | This study |
a
Apr, ampicillin resistance; kan, kanamycin resistance gene; metAW3110, metA gene from E. coli strain W3110.