TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Characteristic(s)a | Reference or source |
|---|---|---|
| Strains | ||
| Lysobacter enzymogenes | ||
| OH11 | Wild type; Kmr | 46 |
| Δ3756 (ΔlchD) | le3756 in-frame deletion mutant | This study |
| Δ2762 (ΔlchP) | lchP in-frame deletion mutant | 27 |
| Δ0082 ΔlchP | le0082 and lchP double-deletion mutant | This study |
| Δ4875 ΔlchP | le4875and lchP double-deletion mutant | This study |
| Δ3882 ΔlchP | le3882 and lchP double-deletion mutant | This study |
| Δ0901 ΔlchP | le0901 and lchP double-deletion mutant | This study |
| Δ2826 ΔlchP | le2826 and lchP double-deletion mutant | This study |
| Δ0155 ΔlchP | le0155 and lchP double-deletion mutant | This study |
| Δ2120 ΔlchP | le2120 and lchP double-deletion mutant | This study |
| Δ3756 ΔlchP | le3756 and lchP double-deletion mutant | This study |
| Δ1158 ΔlchP | le1158 and lchP double-deletion mutant | This study |
| OH11(pBBR) | Vector control | This study |
| OH11(LchD) | Vector-based lchD overexpression strain | This study |
| OH11(LchDGGAAV) | Site-mutated lchD overexpression strain | This study |
| Escherichia coli | ||
| TOP10 | supE44 lacU169(ΔlacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA11 | 47 |
| MG1655 | Wild type | ATCC 700926a |
| XL1-Blue MRF′ | Δ(mcrA 183 83mcrCB-hsdSMR-mrr) 173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac {F[3proAB lacIqZΔM15 Tn10 (Tetr)]} | 43 |
| BL21(DE3) | Host strain for protein expression; Kmr | TaKaRa Company, Shanghai, China |
| Plasmids | ||
| pEX18GM | Suicide vector for gene in-frame deletion; Gmr | 48 |
| 0082-pEX18 | Plasmid for in-frame deletion of le0082 | This study |
| 4875-pEX18 | Plasmid for in-frame deletion of le4875 | 27 |
| 3882-pEX18 | Plasmid for in-frame deletion of le3882 | 27 |
| 0901-pEX18 | Plasmid for in-frame deletion of le0901 | This study |
| 2826-pEX18 | Plasmid for in-frame deletion of le2826 | This study |
| 0155-pEX18 | Plasmid for in-frame deletion of le0155 | 27 |
| 2120-pEX18 | Plasmid for in-frame deletion of le2120 | This study |
| 3756-pEX18 | Plasmid for in-frame deletion of le3756 | This study |
| 1158-pEX18 | Plasmid for in-frame deletion of le1158 | This study |
| pBBR1-MCS5 | Broad-host-range vector for gene overexpression; Gmr | 49 |
| lchD-pBBR | Plasmid for LchD overexpression | This study |
| lchDGGAAV-pBBR | Plasmid for site-mutated LchD overexpression | This study |
| pBAD/Myc-His B | Vector for arabinose-inducible expression; Ampr | Invitrogen |
| GGDEV-pBAD | pBAD/Myc-His B containing the cytoplasmic part of LchD | This study |
| GGAAV-pBAD | pBAD/Myc-His B containing mutated LchD | This study |
| pTRG | Plasmid for protein expression in the bacterial two-hybrid assay; Tetr | 50 |
| LchP-pTRG | pTRG containing the cytoplasmic part of LchP | 27 |
| LchD-pTRG | pTRG containing the cytoplasmic part of LchD | This study |
| PAS-pTRG | pTRG containing the PAS domain of LchP | This study |
| GGDEF-pTRG | pTRG containing the GGDEF domain of LchP | This study |
| EAL-pTRG | pTRG containing the EAL domain of LchP | This study |
| GGAAV-pTRG | pTRG containing site-mutated LchD | This study |
| 1158-pTRG | pTRG containing Le1158 | This study |
| pBT | Plasmid for protein expression in bacterial two-hybridization assays; Chlor | 50 |
| LchP-pBT | pBT containing the cytoplasmic part of LchP | 27 |
| LchD-pBT | pBT containing the cytoplasmic part of LchD | This study |
| pMAL-p2x | Vector for expression of the MBP tag fusion protein | Huayue Company, Beijing, China |
| LchP-pMAL-p2x | pMAL-p2x::LchP PAS+GGDEF+EAL | 27 |
| pET30a(+) | Vector for expression of the His tag fusion protein | TaKaRa Company |
| LchD-pET30a | pET30a::lchD GGDEV | This study |
a
Kmr, Gmr, Ampr, Tetr, and Chlor represent kanamycin, gentamicin, ampicillin, tetracycline, and chloramphenicol resistance, respectively.