TABLE 2.
Plasmids and bacteria in this study
| Strain/plasmid | Relevant feature(s) | Function | Reference or source |
|---|---|---|---|
| E. coli strains | |||
| MG1655 | F−, λ−, rph-1 | Expression, PCR | CGSC, Yale University |
| DH5α | fhuA2Δ(argF-lacZ)U169 phoA glnV44 Φ80 Δ(lacZ)M15 gyrA96 recA1 relA1 endA1 thi-1 hsdR17 | Subcloning | New England Biolabs |
| Plasmids | |||
| pHN1257 | pSC101H ori, Kanr, lacIq, Ptrc, laco-PT-MCS | asRNA expression vector | 5 |
| pHNRF4 | pHN1257 carrying ribF asRNA targeting −50 to +50 of ribF | IPTG-inducible asRNA expression | This study |
| pHNIS | pHN1257 carrying ileS asRNA targeting −59 to +61 of ileS | ||
| pHNLA2 | pHN1257 carrying lspA asRNA targeting −45 to +55 of lspA | ||
| pHNLA3 | pHN1257 carrying lspA asRNA targeting +1 to +100 of lspA | ||
| pHNLA4 | pHN1257 carrying lspA asRNA targeting +21 to +138 of lspA | ||
| pHNLA5 | pHN1257 carrying lspA asRNA targeting +51 to +151 of lspA | ||
| pHNIH | pHN1257 carrying ispH asRNA targeting −49 to +89 of ispH | ||
| p1G2 | pBR322/ColE ori, Cmr, PBAD, AraC N6I, V65G, L133M, E165G, E169V, C280* | ORF expression vector | 23 |
| p1G2RF | p1G2 carrying ribF | l-Arabinose-inducible ORF expression | This study |
| p1G2IS | p1G2 carrying ileS | ||
| p1G2LA | p1G2 carrying lspA | ||
| p1G2IH | p1G2 carrying ispH |