TABLE 1.
Bacterial strains, plasmids, bacteriophages, and primers
| Strain, plasmid, phage, or primer | Relevant characteristic(s)a | Source or reference |
|---|---|---|
| Bacterial strains | ||
| L. lactis subsp. lactis NCK203 | Propagating host for phages | 17, 39 |
| L. lactis subsp. lactis MG1363 | Transformation host | 14 |
| E. coli MC1061 | Transformation host | 18 |
| E. coli TOP10 | Transformation host | Invitrogen |
| Plasmids | ||
| pTRKH2 | E. coli gram-positive cloning vector, 6.7 kb, Emr | 36 |
| pAK80 | E. coli gram-positive cloning and promoter probe vector, 11 kb, Emr | 19 |
| pSMBI 1 (designated pAK80::CI in text) | pAK80 + 907-bp φ31 PCR fragment from primers cl-XhoI-P1 and cI-BamHI-P2 | 28 |
| pTRK676 | pTRKH2 + 907-bp φ31 PCR fragment from primers cI-XhoI-P1 and cI-BamHI-P2. Encodes full-length CI repressor | This study |
| pTRK677 (designated pTRKH2::CI-per1 in text) | Mutated pTRK676 (cI gene with an ochre point mutation) | This study |
| pTRK678 (designated pTRKH2::CI-per2 in text) | pTRKH2 + 571-bp φ31 PCR fragment from primers cI-XhoI-P1 and cI sub 2-BamHI | This study |
| pTRK679 | pTRKH2 + 517-bp φ31 PCR fragment from primers cI-XhoI-P1 and cI sub 3-BamHI | This study |
| pTRK680 | pTRKH2 + 469-bp φ31 PCR fragment from primers cI-XhoI-P1 and cI sub 4-BamHI | This study |
| pTRK681 | pTRKH2 + 424-bp φ31 PCR fragment from primers cI-XhoI-P1 and cI sub 5-BamHI | This study |
| pTRK728 (designated pAK80::CI-per 1 in text) | pAK80 + 907-bp pTRK677 PCR fragment from primers cI-XhoI-P1 and cI-BamHI-P2 | This study |
| pTRK729 (designated pAK80::CI-per2 in text) | pAK80 + 571-bp φ31 PCR fragment from primers cI-XhoI-P1 and cI sub 2-BamHI | This study |
| Phages | ||
| φ31 | P335 group phage | 1 |
| φ31.1 | P335 group phage, recombinant variant of φ31 | 12 |
| φ31.2 | P335 group phage, recombinant variant of φ31 | 12 |
| ul36 | P335 group phage | 33 |
| ul37 | P335 group phage, recombinant variant of ul36 | 33 |
| Q30 | P335 group phage | 32 |
| Q33 | P335 group phage | 32 |
| Q36 | P335 group phage | 32 |
| mm210a | P335 group phage | Rhodia, Inc. |
| mm210b | P335 group phage | Rhodia, Inc. |
| A1 | P335 group phage | Rhodia, Inc. |
| B1 | P335 group phage | Rhodia, Inc. |
| CS | P335 group phage | Rhodia, Inc. |
| D1 | P335 group phage | Rhodia, Inc. |
| φ48 | P335 group phage | Rhodia, Inc. |
| φ50 | P335 group phage | Rhodia, Inc. |
| Primersb | ||
| cI-XhoI-1 (81) | GGC CGC TCG AGC CTG TTC CGT CTG CCG | |
| cI-BamHI-2 | TAG TAG GAT CCT TTT GGG AGA GAT AAA GCG CC | |
| cI sub 2-BamHI (621) | AGC TGG ATC CTT ATT CAG TTT TAA TAG | |
| cI sub 3-BamHI (564) | TAG AGG ATC CTT AAA CAT TTG CAG TAT CTA | |
| cI sub 4-BamHI (516) | TAG AGG ATC CTT ACT TCA TAA CTG TAA TAG | |
| cI sub 5-BamHI (471) | TAG AGG ATC CTT ATG ATT CTC GCG GTT CAA | |
| cI sub 6-BamHI (430) | ATG CGG ATC CTC ACG GCT CAC TAA CT | |
| P1-forward (282) | CGC GGA TCC CAA CCT GTC TTA ATC | |
| P1-reverse (17) | CGC CTG CAG GCT TTT TAT TTG CCA | |
| cI-extend (488) | AAT GAT TGA ACC GCG AGA ATC AG | |
| 75-right (673) | GTA TTG ACC TTG CCG ATT TAG TAG AT | |
| O1 left (241) | CTT TAA TCT CAT TTG AGA GTT T | |
| O1 right (174) | TAT CAT TTG GAA AAT AAT TGT C | |
| O2 left (154) | TGA CAA TTA TTT TCC AAA TGA T | |
| O2 right (99) | GCC GTA CTG AAT GCT CCA TGA T | |
| O3 left | GCT ACA GAA CTT CTT GGT ATT A | |
| O3 right | GTT TTC GTT TTG TGT GAT TGT A | |
| CI NcoI (271) | GAT CCC ATG GCT ATG AGA TTA AAG AAA ATA ATG |
a
Emr, erythromycin resistance.
b
The base pair position for the 3′-terminal nucleotide of primers from the sequence shown in Fig. 1 is shown in parentheses. Underlined portions of sequences represent restriction enzyme recognition sites in primers.