TABLE 1.
Bacterial strains, bacteriophages, and plasmids used in this study
| Strain, phage, or plasmid | Genotype and relevant features | Source or reference |
|---|---|---|
| Bacterial strains | ||
| BL21(DE3) | E. coli B dcm hsdS(rB− mB−) gal(λimm21nin5 int::lacI-lac puv5-T7 gene1) | 43 |
| MC4100 | E. coli K-12 F− araD139 Δ(argF-lac)U169 rpsL150 relA1 flbB5301 deoC1 ptsF25 rbsR | 37 |
| XL1-Blue | E. coli K-12 recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F′ proAB lacIqZΔM15::Tn10 (Tetr)] | Stratagene |
| Phages | ||
| λS+ | cI857 | Laboratory stock |
| λ Cmr Δ(SR) | stf::cat::tfa cI857 Δ(SR); replacement of stf and tfa genes (λ nt 19996–22220) with cat gene (36); Δ(SR); loss of λ nt 45136–45815 (31) | Laboratory stock |
| Plasmids | ||
| pOR19 | Derivative of pBR322; EcoRI site inactivated by blunt ending and ligation | 34 |
| pKB1 | Derivative of pOR19; λ lysis gene region (Sam7) cloned as HindIII/ClaI fragment (λ nt 44141–46440) | 26a |
| pKB110 | Derivative of pOR19; λ lysis gene region cloned as HindIII/ClaI fragment (λ nt 44141–46440) | 9 |
| pSg38p | Site-directed mutagenesis derivative of pKB110; S Gly38 mutated to Pro to create a SmaI site in codons 38 and 39 | This work |
| pET11a | Expression vector containing hybrid T7-lacPO promoter, φ10 ribosome binding site, and lacI gene (pBR322 derivative) | Novagen |
| pETNH | N-terminal oligohistidine tag (MH6GSH)-coding region inserted into pET11a as NdeI/BglII fragment | This study |
| pETNHS105 | Same as pETNHS107, but encodes NHS105 | This study |
| pS105 | Seamless Cloning derivative of pSg38p; Met1Leu mutation; encodes S105 only | This study |
| pNHS105 | Seamless Cloning derivative of pSg38p; oligohistidine tag inserted at the N terminus of S105 | This study |
| pCHS105 | Seamless Cloning derivative of pSg38p; oligohistidine tag inserted at the C terminus of S105 | This study |
| pS105H94 | Modified Seamless Cloning derivative of pS105; oligohistidine tag (G2H6G2) inserted behind Phe94 | This study |
| pS105H94a52v | QuikChange mutagenesis derivative of pS105H94 with Ala52Val allele | This study |
| pETS105H94 | Derivative of pET11a; S105H94 allele inserted as Nde/BamHI fragment | This study |
| pETS105H94a52v | Derivative of pET11a; S105H94a52v allele inserted as NdeI/BamHI fragment | This study |