TABLE 1.
Plasmids used in this study
| Plasmid | Description and/or source (reference) |
|---|---|
| pMRC01 | 60.2-kb natural lactococcal plasmid containing ltnEFIRA1A2M1TM2J (12) |
| pMRC01S7G | Derivative of pMRC01 generated by site-directed mutagenesis (10) |
| pCI372 | High-copy-number E. coli-Lactococcus cloning vector (14) |
| pOM02 | pCI372-ltnEFIRA1A2M1TM2J (21) |
| pOM18 | Equivalent to pOM02ΔltnM1 (20) |
| pOM31 | Equivalent to pOM02ΔltnA2 (20) |
| pOM32 | Equivalent to pOM02ΔltnM2 (20) |
| pOM39 | Equivalent to pOM02ΔltnA1 (20) |
| pOM11 | 6-kb fragment from pOM02 (primers 5145 and 5146) cloned into BamHI/SalI-digested pCI372 |
| pOM27 | pCI372-ltnEFIR; 4-kb PCR product (primers 5144 and 5558) cloned into KpnI/BamHI-digested pOM11 |
| pOM25 | Equivalent to pOM02ΔltnFE; 4.3-kb product (primers 5144 and 5556) cloned into SacI/BamHI site of pOM11 |
| pOM28 | Equivalent to pOM02ΔltnJ; 4.7-kb product [primers LtnX(f) and LtnX(r)] cloned into BamHI/SalI-digested pOM02 |
| pOM34 | Equivalent to pOM02ΔltnR; 2.4-kb product [primers FP(OP)372 and 5557] cloned into SacI/KpnI-digested pOM27 |
| pOM38 | 4-kb product (primers 6163 and 5144) cloned into KpnI/BamHI-digested pOM11 |
| pOM40 | Equivalent to pOM02ΔltnI; 2-kb product [primers FP(OP)372 and 6164] cloned into SacI/KpnI-digested pOM38 |
| pOM44 | Equivalent to pOM02ΔltnA1A2; replacement of a 3.5-kb SacI/KpnI fragment from pOM31 with a 2.9-kb SacI/KpnI fragment from pOM39 |
| pOM51 | Equivalent to pOM02ΔltnM1M2; replacement of a BamHI/SacI fragment from pOM18 with the corresponding region from pOM32 |
| pPCM2J | 4.7-kb product (primers ΔT for and pOM02rev) cloned into BamHI/SalI-digested pCI372 |
| pPCΔT | Equivalent to pOM02ΔltnT; 6-kb product [primers 5144 and FP(OP)372] cloned into BamHI/SacI-digested pPCM2J |
| pDF01 | pCI372-ltnA1A2; 1-kb product (primers A1soeA and A2soeD) cloned into PstI/EcoRI-digested pCI372 |