TABLE 1.
Strains and plasmids used in this work
| Strain or plasmid | Genotype or descriptiond | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| PERM590 | E. coli XL10-Gold carrying plasmid pPERM590 (Ampr Kmr) | 8 |
| PERM594 | E. coli XL10-Gold carrying plasmid pPERM594 (Ampr Kmr) | 8 |
| B. subtilis | ||
| PS832 | Wild-type; trp+ revertant of strain 168 | Laboratory stock |
| YB955 | hisC952 metB5 leuC427 xin-1 SpβSENS | 42 |
| PERM563 | ΔytkD::neo Neor | pPERM590→YB955b |
| PERM564 | ΔmutT::tet Tetr | pPERM594→YB955b |
| PERM597 | ΔytkD::neo ΔmutT::tet Neor Tetr | pPERM594→PERM563b |
| PERM598c | ΔyfhQ::spc Spcr | P. Setlow |
| PERM599c | ΔmutM::tet Tetr | P. Setlow |
| PERM571 | ΔmutM::tet Tetr | PS599→YB955a |
| PERM572 | ΔmutM::tet ΔyfhQ::spc Spcr Tetr | PERM599→PERM571a |
| PERM573 | ΔytkD::neo DmutM::tet ΔyfhQ::spc Neor Spcr Tetr | pPERM590→PERM572a |
| PERM731 | ΔytkD::neo DmutM::tet ΔyfhQ::spc with a Pspac-mutSL-lacI construct from pMPR002 inserted into the amyE locus; Neor Spcr Tetr Cmr | pMPR002→PERM573a |
| Plasmids | ||
| pDG148 | spac expression vector; Ampr Kmr | W. L. Nicholson |
| pDG364 | Integration vector (promotes ectopic integration into the amyE locus); Ampr Cmr | 9 |
| pPERM590 | pBEST501 containing 798-bp HindIII-SalI PCR fragment encompassing 589 bp upstream and 209 bp downstream of the ytkD translational start codon and an 866-bp BamHI-SacI PCR fragment encompassing 131 bp upstream and 735 bp downstream of the ytkD translational stop codon; Ampr | 8 |
| pPERM594 | PDG1515 containing the 986-bp BamHI-XbaI fragment encompassing 904 bp upstream and 82 bp downstream of the mutT translational start codon and 980-bp EcoRI-HindIII fragment encompassing 193 bp upstream and 787 bp downstream of the mutT translational ending codon; Ampr | 8 |
| pMPR001 | ∼4.5-kb B. subtilis mutSL open reading frame cloned into the SalI/XbaI sites of pDG148 | This study |
| pMPR002 | pDG364 with an ∼6.28-kb EcoRI/BamHI fragment from pMPR001 containing the Pspac-mutSL-lacI construct | This study |
a
Chromosomal DNA from the strain to the left of the arrow was used to transform the strain to the right.
b
Plasmid DNA from the strain to the left of the arrow was used to transform the strain to the right.
c
The background for this strain is PS832.
d
Amp, ampicillin; Km, kanamycin; Neo, neomycin; Tet, tetracycline; Spc, spectinomycin; Cm, chloramphenicol.