TABLE 1.
Bacterial strains, plasmids, phages, and primers used in this study
| Strain, plasmid, bacteriophage, or primer | Genotype/relevant features | Source or reference |
|---|---|---|
| Strains | ||
| E. coli K-12 | ||
| P90A5c | Prototrophic; parent of 397c | Christie et al. (10) |
| 397c | Ts; carries rpoC397 deletion | Christie et al. (10) |
| SA1030 | his strA gal-3 | Adhya and Shapiro (1) |
| AD1600 | his strA gal-3 rho(Ts)15 | Das et al. (13) |
| DH5α | Auxotrophic rK− mK+ | BRL Life Technologies |
| MC4100 | araD139Δ(argF-lac)U169 thiA1 rpsL150 relA1 deoC1 PtsF25 rbsR flbB5301 | Zhang and Young (43) |
| RW4206 | Met+rpoC+ transductant of MG1655 metA | Robert Weisberg |
| RW4204 | Met+rpoC397 transductant of MG1655 metA | Robert Weisberg |
| E. coli C | ||
| C-1a | Prototrophic F− | Sasaki and Bertani (32) |
| C-520 | supD F+; P2 indicator | Sunshine et al. (36) |
| C-4508 | Auxotrophic F− | King et al. (20) |
| C-4517 | C4508 supD | King et al. (20) |
| C-4518 | C4508 supF | King et al. (20) |
| Plasmids | ||
| pCYB2 | Expression vector | New England Biolabs |
| pCYB2lysB+ | This work | |
| pCYB2lysC+ | This work | |
| pCYB2lysC(trl) | This work | |
| pCYB2lysC(gor) | This work | |
| pCYB2lysB+lysC+ | This work | |
| pCYB2lysB+lysC(gor) | This work | |
| pCYB2β′ | rpoC+ expression vector | Nedea et al. (28) |
| pCYB2β′397c | rpoC397 expression vector | This work |
| pBBRV7 | P2 EcoRV fragment (6290-8378) in pUC18/HincII | Ziermann et al. (44) |
| pDEB50 | Delta expression vector with modified T7A1 (PA1/04/03) promoter and lacIq | Christie et al. (9) |
| pFCAT100 | P2 pF-cat expression plasmid | Grambow et al. (15) |
| pGC160 | pUCF4 with wild-type P2 lysis cassette | This work |
| pGC163 | pUCF4 with P2 gor lysis cassette | This work |
| pGCRV7gor-1 | P2 gor-1 EcoRV fragment (6290-8378) in pUC18/HincII | This work |
| pNL130 | P2 BstEII fragment (7455-8862) in pUC9/HincII | Linderoth et al. (24) |
| pSL130 | Ptac cat expression plasmid | Li et al. (22) |
| pTG257 | Ogr expression vector with modified T7A1 (PA1/04/03) promoter and lacIq | This work |
| pTG605 | pNL130 with lysC(Am) mutation | This work |
| pUC18 | Cloning vector | Norrander et al. (29) |
| pUCF4 | pUC8 carrying P2 pF promoter | Christie and Calendar (12) |
| pRG1 | pACYC177/PstI with 1.2-kb PstI fragment carrying lacIq | Robert Garcea |
| Bacteriophages | ||
| P2 vir-22 | Immunity insensitive | Chattoraj and Inman (8) |
| P2 vir-22 gor-1 | Spontaneous mutant; plates on AD1600, 397c | This work |
| P2 vir-22 trl-21 | Spontaneous mutant, plates on 397c | This work |
| P2 vir-22 trl-22 | Spontaneous mutant, plates on 397c | This work |
| P2 vir-1 | Clear plaque mutant | Bertani (5) |
| P2 vir-1 R(Am)3 | Tail assembly defect | Lindahl (23) |
| P2 vir-1 lysC(Am) | This work | |
| 186 c(Ts) | Baldwin et al. (2) | |
| Hy5 | P2/186 hybrid | Hocking and Egan (17) |
| λ | stf::cat::tfa cI857 | Zhang and Young (43) |
| λRz(Am) Rz1+ | Zhang and Young (43) | |
| λRz+Rz1(Am) | Zhang and Young (43) | |
| λRz(Am) Rz1(Am) | Zhang and Young (43) | |
| Primersa | ||
| MR2 | GCACGCCAGTGATTGTC | Linderoth et al. (24) |
| 28am | ACGGCGATTTATAGGCCGATATCCGG | This work |
| lysB_Up | CATATGTCAAGGCTGATGATTG | This work |
| lysC_Up | CATATGAGAACGAAGATTTTCG | This work |
| lysC_Down | CTCGAGTCAGTCAGCGCCCTGCGCA | This work |
| Lys3 | GAACTGCAGTCAATCCACGAATATCCGCAG | This work |
| Lys5 | CGAGGATCCGTCAATCTGTGGGAGTAAC | This work |
a
Primer sequences are given 5′→3′.