TABLE 1.
Strains and plasmids
| Strain or plasmid | Relevant genotype and/or characteristic(s) | Source or reference |
|---|---|---|
| Strains | ||
| S. salivarius | ||
| ATCC 25975 | Wild-type, Lac+ Glu+ Gal+ | I. R. Hamilton, University of Manitoba |
| G71 | El-negative mutant derived from S. salivarius ATCC 25975 | 9 |
| E. coli | ||
| BL21 (DE3) | F−ompT hsdSB(rB− mB−) gal dcm (DE3) | Novagen |
| XL1-Blue | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac[F′ proAB lacIqZΔM15 Tn10 (Tetr)] | Stratagene |
| LMG194 | F− ΔlacX74 galE thi rpsL ΔphoA (PvuII) Δara714 leu::Tn10 | Invitrogen |
| TOP10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 deoR recA1 araD139 Δ(ara-leu)7697 galU rpsL endA1 nupG | Invitrogen |
| Plasmids | ||
| pET-28a(+) | Expression vector | Novagen |
| pET-29a(+) | Expression vector | Novagen |
| pET-19b | Expression vector | Novagen |
| pBAD/His | Expression vector | Invitrogen |
| pCR-Blunt | Cloning vector | Invitrogen |
| pETI-16 | Contains the ptsI gene of S. salivarius cloned into pET-28a(+) | This work |
| pLacSIIA | Contains the last 519 nucleotides of lacS from S. salivarius (which code for the IIA domain), cloned into pET29a(+) | This work |
| pLacSIIAH552R | Contains the last 519 nucleotides of lacS from S. salivarius (which code for the IIA domain), with a mutation replacing LacS H552 by R, cloned into pET29a(+) | This work |
| pTML2 | Contains the manL gene of S. salivarius cloned into pET19b | This work |
| pDR3 | Contains the manH gene of S. salivarius cloned into pET19b | This work |
| pHPW18 | Contains the ptsH gene of S. salivarius cloned into pBAD | 8 |
| pHPK229 | Contains the hprK gene of S. salivarius cloned into pCR-Blunt | 3 |