Table 1.
Strains and plasmids used in this study
| F. johnsoniae strain or plasmid | Genotype and/or descriptiona | Reference(s) or source |
|---|---|---|
| Strains | ||
| UW101 (ATCC 17061) | Wild type | 38, 39 |
| UW102-57 | Spontaneous gldK mutant | 17, 26 |
| UW102-141 | Nitrosoguanidine-induced gldK mutant | 17, 26 |
| CJ1372 | gldK::HimarEm1 (Emr) | 17 |
| CJ1373 | gldK::HimarEm1 (Emr) | 17 |
| CJ1827 | rpsL2 (Smr); wild-type strain used in construction of deletion mutants | 20 |
| CJ1922 | rpsL2 ΔsprB (Smr) | 20 |
| CJ1984 | rpsL2 ΔremA (Smr) | 4 |
| CJ1985 | rpsL2 ΔsprB ΔremA (Smr) | 4 |
| CJ2083 | rpsL2 remA::myc-tag-1 (Smr) | 4 |
| CJ2090 | rpsL2 Δ(gldN gldO) (Smr) | 4 |
| CJ2122 | rpsL2 ΔgldK (Smr) | This study |
| CJ2140 | rpsL2 ΔgldK in remA::myc-tag-1 background (Smr) | This study |
| CJ2157 | rpsL2 ΔgldL (Smr) | This study |
| CJ2262 | rpsL2 ΔgldM (Smr) | This study |
| CJ2263 | rpsL2 ΔgldM in remA::myc-tag-1 background (Smr) | This study |
| CJ2281 | rpsL2 ΔgldL in remA::myc-tag-1 background (Smr) | This study |
| CJ2089 | rpsL2 Δ(gldN gldO) remA::myc-tag-1 (Smr) | 4 |
| CJ2302 | rpsL2 ΔsprA (Smr) | This study |
| CJ2317 | rpsL2 ΔsprA in remA::myc-tag-1 background (Smr) | This study |
| CJ2327 | sprT disruption in remA::myc-tag-1 background (Emr Smr) | This study |
| KDF001 | sprT mutant (Emr) | 7 |
| FJ149 | sprE mutant (Emr) | 6 |
| Plasmids | ||
| pET30a | Protein expression vector; Kmr | Novagen |
| pCP23 | E. coli-F. johnsoniae shuttle plasmid; Apr (Tcr) | 22 |
| pCP29 | E. coli-F. johnsoniae shuttle plasmid; Apr (Cfr Emr) | 24 |
| pRR51 | rpsL-containing suicide vector; Apr (Emr) | 20 |
| pJVB4 | 1.3-kbp fragment of gldK amplified with primer pair 704/705 and ligated into EcoRI-, SalI-digested pET30a; Kmr | This study |
| pJVB2 | 0.6-kbp fragment of gldL amplified with primer pair 706/707 and ligated into EcoRI-, SalI-digested pET30a; Kmr | This study |
| pJVB6 | 1.3-kbp fragment of gldM amplified with primer pair 708/709 and ligated into EcoRI-, SalI-digested pET30a; Kmr | This study |
| pAB18 | 2.4-kbp region downstream of gldL amplified with primers 1199 and 1200 and ligated into XbaI-, SalI-digested pRR51; Apr (Emr) | This study |
| pAB19 | Construct used to delete gldL; 2.2-kbp region upstream of gldL amplified with primer pair 1197/1198 and ligated into BamHI-, XbaI-digested pAB18; Apr (Emr) | This study |
| pAB30 | Construct used to delete sprA; 2.5 kbp upstream and 2.5 kbp downstream of sprA amplified using primer pairs 1333/1334 and 1335/1336, respectively, and ligated into BamHI-, SalI-digested pRR51; Apr (Emr) | This study |
| pJJ01 | Construct used to delete gldK; 1.9-kbp region upstream and 1.9-kbp region downstream of gldK amplified using primer pairs 1209/1210 and 1211/1212, respectively, and ligated into BamHI-, SphI-digested pRR51; Apr (Emr) | This study |
| pJJ02 | Construct used to delete gldM; 2.6-kbp region upstream and 2.4-kbp region downstream of gldM amplified using primer pairs 1237/1214 and 1215/1238, respectively, and ligated into BamHI-, SphI-digested pRR51; Apr (Emr) | This study |
| pTB99 | pCP23 carrying gldK; Apr (Tcr) | 17 |
| pTB81a | pCP23 carrying gldL; Apr (Tcr) | 17 |
| pTB94a | pCP23 carrying gldM; Apr (Tcr) | 17 |
| pTB79 | pCP23 carrying gldN; Apr (Tcr) | 17 |
| pKF002 | pCP23 carrying sprT; Apr (Tcr) | 7 |
| pSN48 | pCP23 carrying sprA; Apr (Tcr) | 18 |
a
Antibiotic resistance phenotypes are as follows: ampicillin, Apr; cefoxitin, Cfr; erythromycin, Emr; streptomycin, Smr; tetracycline, Tcr. Unless indicated otherwise, the antibiotic resistance phenotypes are those expressed in E. coli. The antibiotic resistance phenotypes given in parentheses are those expressed in F. johnsoniae but not in E. coli.