TABLE 2.
Bacterial strains and plasmids used in this study
| Strain, plasmid, or primer | Characteristic(s)a | Source or reference |
|---|---|---|
| Strains | ||
| S. thermophilus | ||
| LMD-9 | Wild type | 15 |
| LF105 (Δ1) | LMD-9 Δ(blpDSt-blpFSt) | 15 |
| LF106 (Δ7) | LMD-9 Δ(blpGSt-blpXSt) | 15 |
| LF109 (Δ2) | LMD-9 Δ(blpUSt-blpFSt) | This study |
| LF110 (Δ3) | LMD-9 Δ(blpDSt-orf6) | This study |
| LF109.1 (Δ9) | LMD-9 Δ(blpUSt-blpFSt) ΔblpDSt | This study |
| LF109.2 (Δ10) | LMD-9 Δ(blpUSt-blpFSt) Δ(blpDSt-orf1) | This study |
| LF111 (Δ4) | LMD-9 Δ(blpDSt-orf2) | This study |
| LF111.1 (Δ4/Δ5) | LMD-9 Δ(blpDSt-orf2) Δ(blpESt-blpFSt) | This study |
| LF112 (Δ5) | LMD-9 Δ(blpESt-blpFSt) | This study |
| LF113 (Δ6) | LMD-9 Δ(blpUSt-orf6) | This study |
| LF114 (Δ8) | LMD-9 ΔblpGSt | This study |
| L. plantarum | ||
| NCIMB8826 | Wild type | NCIMBb |
| L. lactis | ||
| NZ3900 | MG1363 derivative | 7 |
| E. coli | ||
| TG1 | supE hsdΔ5 thi Δ(lac-proAB) F′[traD36 proAB+lacIqlacZΔM15] | 38 |
| EC1000 | Kmr RepA+ | 26 |
| AH50 | MC1000 phoR Δara714 leu+phoA68 | 21 |
| AH55 | AH50 dsbA::kan-1 | 21 |
| Plasmids | ||
| pGIBG001 | Emr; pMG36e in which an NcoI restriction site has been introduced to allow the cloning of genes in translational fusion with the P32 expression cassette | 15 |
| pGILF003 | Emr; pGIBG001 with a 0.69-kb insert containing the blpGSt ORF of S. thermophilus LMD-9 in translational fusion with the P32 expression cassette | This study |
| pMG36e | Emr; E. coli-S. thermophilus shuttle vector; contains the P32 promoter | 40 |
| pMG36eT | Emr; pMG36e with a 0.36-kb insert containing the 3′ end of the ldhL ORF and the transcriptional terminator of ldhL from L. plantarum WCFS1 | This study |
| pGILF004 | Emr; pMG36eT with a 0.6-kb insert containing the blpDSt-orf2 operon of S. thermophilus LMD-9 in transcriptional fusion with the P32 promoter | This study |
| pGILF005 | Emr; pMG36eT with a 0.6-kb insert containing the blpDSt-orf2 operon and a 3.55-kb insert containing the blpABSt ORFs of S. thermophilus LMD-9 in transcriptional fusion with the P32 promoter | This study |
| pGhost9 | Emr Ts | 28 |
| pGILF109 | Emr Ts; pGhost9 derivative for the deletion of blpUSt-blpFSt | This study |
| pGILF109.1 | Emr Ts; pGhost9 derivative for the in-frame deletion of blpDSt in strain LF109 | This study |
| pGILF109.2 | Emr Ts; pGhost9 derivative for the deletion of blpDSt-orf1 in strain LF109 | This study |
| pGILF110 | Emr Ts; pGhost9 derivative for the deletion of blpDSt-orf6 | This study |
| pGILF111 | Emr Ts; pGhost9 derivative for the deletion of blpDSt-orf2 | This study |
| pGILF112 | Emr Ts; pGhost9 derivative for the deletion of blpESt-blpFSt | This study |
| pGILF113 | Emr Ts; pGhost9 derivative for the deletion of blpUSt-orf6 | This study |
| pGILF114 | Emr Ts; pGhost9 derivative for the deletion of blpGSt | This study |
a
Kmr, Emr, and Ts indicate resistance to kanamycin, resistance to erythromycin, and temperature-sensitive mutations in the RepA replication protein, respectively.
b
NCIMB, The National Collections of Industrial and Marine Bacteria, Ltd., Aberdeen, Scotland.