TABLE 1.
List of bacteria, phages, and plasmids used in the study
| Bacterial strain, phage, or plasmid | Relevant characteristic(s)a | Source or reference |
|---|---|---|
| L. lactis strains | ||
| IL1403 | L. lactis subsp. lactis IL1403; host for phages P008, bIL170, and bIL67 | 7 |
| MB112 | L. lactis subsp. cremoris MG1363 Δupp; host for phages p2, sk1, jj50, and c2 | 50 |
| NZ9000 | L. lactis subsp. cremoris MG1363 pepN::nisRK | 40 |
| JH-20 | MB112(pJH2) Camr Abi+ | 35 |
| JH-22 | IL1403(pJH2) Camr Abi+ | 35 |
| JH-32 | MB112 containing pGhost9::ISS1 insert upstream of abiV on chromosome; Ermr Abi+ | 35 |
| JH-74 | NZ9000(pJH14) Camr | This study |
| JH-76 | NZ9000(pNZ8010) Camr | This study |
| JH-86 | MB112(pJH13, pAJ80) Camr Tetr | This study |
| JH-87 | MB112(pMAP84, pAJ80) Camr Tetr | This study |
| E. coli strains | ||
| MC1000 | Laboratory strain; cloning host | 64 |
| DH5α | Laboratory strain; cloning host | 70 |
| AJ177 | DH5α(pAJ80) Camr | DTUb strain collection |
| M15 | (pREP4) Kmr; used for cloning in pQE-70 His tag vector | Qiagen |
| JH-60 | AJ177(pJH13) Camr Tetr | This study |
| JH-65 | M15(pJH12) Ampr Kmr | This study |
| JH-77 | M15(pQE-70) Ampr Kmr | This study |
| JH-84 | DH5α(pJH15) Tetr | This study |
| JH-88 | AJ177(pJH15) Tetr | This study |
| JH-89 | MC1000(pJH14) Camr | This study |
| Phages | ||
| p2 | Small isometric headed; 936 species | 56 |
| sk1 | Small isometric headed; 936 species | 14 |
| jj50 | Small isometric headed; 936 species | 48 |
| P008 | Small isometric headed; 936 species | 48 |
| bIL170 | Small isometric headed; 936 species | 20 |
| c2 | Prolate headed; c2 species | 45 |
| bIL67 | Prolate headed; c2 species | 73 |
| p2.1 | Small isometric headed; 936 species; carries deletion and point mutation in orf26 | This study |
| p2.2, p2.5, p2.7, p2.8 | Small isometric headed; 936 species; carry nonsense mutation in orf26 | This study |
| p2.4 | Small isometric headed; 936 species; carries point mutation in orf26 start codon | This study |
| p2.3, p2.9, p2.10, p2.11 | Small isometric headed; 936 species; carry point mutation in RBS of orf26 | This study |
| p2.6 | Small isometric headed; 936 species; carries nonsense mutations in orf26 and orf28 | This study |
| sk1.1 | Small isometric headed; 936 species; carries mutation in orf26 start codon | This study |
| jj50.1 | Small isometric headed; 936 species; carries nonsense mutation in orf25 (homologue of p2 orf26) | This study |
| P008.1 | Small isometric headed; 936 species; carries nonsense mutation in orf33 (homologue of p2 orf26) | This study |
| bIL170.1 | Small isometric headed; 936 species; carries nonsense mutation in e24 (homologue of p2 orf26) | This study |
| c2.1 | Prolate headed; c2 species; carries mutation in e11 (homologue of p2 orf26) | This study |
| bIL67.1 | Prolate headed; c2 species; carries mutation in orf8 (homologue of p2 orf26) | This study |
| Plasmids | ||
| pAB223 | Vector containing genetic switch of phage TP901-1 | 12 |
| pAJ80 | pCI372 with cloned CI repressor gene from phage TP901-1 | 46 |
| pJH2 | Shuttle expression vector pLC5 with cloned abiV gene; Camr | 35 |
| pJH12 | pQE-70 with cloned fragment comprising nt 1005 to 1388; Ampr | This study |
| pJH13 | pMAP84 with cloned fragment comprising nt 872 to 1446; Tetr | This study |
| pJH14 | pNZ8010 with cloned fragment comprising nt 872 to 1446; Camr | This study |
| pJH15 | pMAP84 with cloned fragment from p2.1 corresponding to nt 872 to 1446; Camr | This study |
| pMAP84 | pPTPL with cloned PL promoter of phage TP901-1 (Fig. 1); Tetr | This study |
| pNZ8010 | PnisA-gusA cat Camr | 23 |
| pPTPL | Low-copy-number E. coli-L. lactis shuttle and promoter-cloning vector; Tetr | 63 |
| pQE-70 | Cloning vector for C-terminal His tagging of proteins; Ampr | Qiagen |
a
The fragments referred to are fragments of the sequence determined in this study. Abi+, phage resistance phenotype; Ampr, ampicillin resistance; Camr, chloramphenicol resistance; Ermr, erythromycin resistance; Kmr, kanamycin resistance; Tetr, tetracycline resistance.
b
DTU, Technical University of Denmark.