TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype and/or descriptiona | Source or reference |
|---|---|---|
| F. johnsoniae strains | ||
| ATCC 17061 strain UW101 | Wild type | 3, 26 |
| UW102-3 | Spontaneous sprA mutant | 19, 62 |
| UW102-57 | Spontaneous gldK mutant | 4, 63 |
| UW102-176 | Nitrosoguanidine-induced gldM mutant | 4, 19 |
| UW102-344 | Spontaneous gldL mutant | 19, 62 |
| CJ1631A | Δ(gldN gldO) | 19 |
| CJ1808 | chiA disruption mutant; (Emr) | This study |
| CJ1827 | Strain used for construction of deletion mutants; rpsL2 (Smr) | 30 |
| CJ2325 | Mutant lacking CTD-encoding region of chiA; rpsL2 (Smr) | This study |
| CJ2355 | Fjoh_4175 deletion mutant; rpsL2 (Smr) | This study |
| FJ149 | sprE disruption mutant; (Emr) | 20 |
| KDF001 | sprT disruption mutant; (Emr) | 6 |
| Plasmids | ||
| pCP23 | E. coli-F. johnsoniae shuttle plasmid; Apr (Tcr) | 64 |
| pET30a | Protein expression vector; Kmr | Novagen |
| pKF002 | pCP23 carrying sprT; Apr (Tcr) | 6 |
| pLYL03 | Suicide vector used for insertional mutagenesis; Apr (Emr) | 65 |
| pME-mCherry | Plasmid expressing fluorescent protein mCherry; Kmr | 66 |
| pNap2 | pCP23 carrying sprE; Apr (Tcr) | 20 |
| pRR51 | rpsL-containing suicide vector for construction of deletions; Apr (Emr) | 30 |
| pSAM1 | 1,299-bp fragment of chiA in pLYL03 for constructing chiA disruption mutant CJ1808; Apr (Emr) | This study |
| pSN48 | pCP23 carrying sprA; Apr (Tcr) | 67 |
| pSSK05 | pCP23 carrying chiA; Apr (Tcr) | This study |
| pSSK07 | 1,833-bp fragment of chiA inserted into pET30a; Kmr | This study |
| pSSK26 | 2,121-bp fragment upstream of chiA CTD-encoding region in pRR51; Apr (Emr) | This study |
| pSSK27 | Construct used to delete CTD-encoding region of chiA; 2,033-bp region downstream of chiA inserted into pSSK26; Apr (Emr) | This study |
| pSSK30 | pCP23 carrying mcherry; Apr (Tcr) | This study |
| pSSK32 | 2,118-bp region downstream of Fjoh_4175 inserted into pRR51; Apr (Emr) | This study |
| pSSK34 | Construct used to delete Fjoh_4175; 1,948-bp region upstream of Fjoh_4175 inserted into pSSK32; Apr (Emr) | This study |
| pSSK45 | mcherry with stop codon amplified with primers 862 and 1443 and cloned into pCP23; Apr (Tcr) | This study |
| pSSK51 | 484-bp fragment spanning the chiA promoter, start codon, and N-terminal signal peptide-encoding region inserted into pSSK30; Apr (Tcr) | This study |
| pSSK52 | 566-bp region encoding 105-amino-acid CTDChiA inserted into pSSK51; Apr (Tcr) | This study |
| pSSK54 | 484-bp fragment spanning the chiA promoter, start codon, and N-terminal signal peptide-encoding region inserted into pSSK45; Apr (Tcr) | This study |
| pTB79 | pCP23 carrying gldN; Apr (Tcr) | 63 |
| pTB81a | pCP23 carrying gldL; Apr (Tcr) | 63 |
| pTB94a | pCP23 carrying gldM; Apr (Tcr) | 63 |
| pTB99 | pCP23 carrying gldK; Apr (Tcr) | 63 |
a
Antibiotic resistance phenotypes are as follows: ampicillin, Apr; erythromycin, Emr; kanamycin, Kmr; streptomycin, Smr; and tetracycline, Tcr. The antibiotic resistance phenotypes given in parentheses are those expressed in F. johnsoniae but not in E. coli. The antibiotic resistance phenotypes without parentheses are those expressed in E. coli but not in F. johnsoniae.