Table 1.
Bacterial strains and plasmids used in this study.
| Strain or plasmid | Relevant characteristics | Reference or source |
|---|---|---|
| Strains | ||
| Escherichia coli BL21 (DE3) | F-, ompI, hsdSB (r-B m-B) gal, dam, met | Jeong et al., 2009 |
| E. coli DH5α | supE44 lacU169 (∅80lacZΔM15) hsdR17 (rk-mk-), recA1 endA1 gyrA96 thi-1 relA1 | Woodcock et al., 1989 |
| E. coli HB101 | F- Δ(gpt-proA)62 leuB6 supE44 ara-14 galK2 lacY1Δ(mcrC-mrr) rpsL20 (Smr) xyl-5 mtl-1 recA13 thi-1 | Boyer and Roulland-Dussoix, 1969;Kessler et al., 1992 |
| E. coli CC118λpir | Rifr; Δ(ara-leu) araDΔlacX74 galE galK phoA20 thi-1 rpsE rpoB argE(Am) recA1 Tn7 λpir | Herrero et al., 1990 |
| Pseudomonas aeruginosa PAO1 | Reference strain | Stover et al., 2000 |
| P. aeruginosa PAO1-Km | Kmr; wild type PAO1 with a Km cassette inserted in a neutral position downstream of glmS | This study |
| P. aeruginosa PAO1 ΔmcpK | P. aeruginosa PAO1 deletion mutant of ΔmcpK gene | This study |
| Plasmids | ||
| pGEM-T | Apr; TA-cloning vector | Promega |
| pET28b(+) | Kmr; Protein expression plasmid | Novagen |
| p34S-Km3 | Kmr, Apr; Km3 antibiotic cassette | Dennis and Zylstra, 1998 |
| pET28-McpK-LBD | Kmr; pET28b(+) derivative containing DNA fragment encoding McpK-LBD | This study |
| pUC18NotI | Apr; ori pMB1, similar to pUC18 but with NotI sites flanking the MCS; cloning vector | Purchased from Biomedal |
| pUC18NotI-5072Up | Apr; pUC18NotI derivative containing a 0.3-kb HindIII-XbaI fragment of upsteam region of pa5072 | This study |
| pMAMV257 | Apr; 1.4-kb PCR product containing the intergenic region between genes pa5548 and pa5549 (glmS) of P. aeruginosa PAO1 was inserted into the EcoRI/HindIII sites of pUC18NotI. A BamHI site was inserted into this intergenic region by PCR. | This study |
| pMAMV258 | Apr, Kmr; 0.96-kb BamHI fragment containing km3 cassette of p34S-Km3 was inserted into the BamHI downstream of glmS in pMAMV257 | This study |
| pUC18NotI-5072UpDw | Apr; pUC18NotI derivative containing a 1-kb HindIII-EcoRI fragment containing upstream and downstream region of pa5072 | This study |
| pKNG101 | Smr; oriR6K, mob, sac; suicide vector | Kaniga et al., 1991 |
| pKNG101-PA5072UpDw | Smr; pKNG101derivative containing a 0.6 Kb NotI fragment from pUC18NotI-5072UpDw cloned into pKNG101 | This study |
| pMAMV261 | Smr, Kmr; 2.4-kb NotI fragment of pMAMV258 was cloned at the same site in pKNG101 | This study |
| pRK600 | Cmr; oriColE1, RK2 mob+, tra+; helper plasmid | de Lorenzo et al., 1990 |
| pBBR1MCS-2 | Kmr; broad-host-range cloning plasmid pBB1MCS derivative containing Kmr cassette, mob+, rep+ | Kovach et al., 1995 |
| pBBR1MCS-2-McpK | Kmr; pBBR1MCS-2 derivative containing mcpK gene and its 607 bp upstream region | This study |
The following abbreviations were used for antibiotics: km, kanamycin; cm, chloramphenicol; ap, ampicillin; rif, rifampicin; sm, streptomycin.