TABLE 1.
Strains and plasmids used in this study
| Features | Reference | |
|---|---|---|
| Strains | ||
| P. putida KT2440 | mt-2 pWW0 cured, TolS− | (62) |
| P. putida DOT-T1E | Tol+, wild type | (63) |
| P. putida KT2440R | Rifr, derivative of P. putida KT2440 | (64) |
| P. putida KT2440R-TK1038 | Rifr, cheR1::mini-Tn5 Kmr | (48) |
| P. putida KT2440R-TK1116 | KT2440R with plasmid pCHESI-3760 inserted into the cheR3 gene, Kmr (KT2440R PP3760::ΩKm, Kmr) | This study |
| P. putida KT2440R-TK1117 | KT2440R with plasmid pCHESI-4392 inserted into the cheR2 gene, Kmr (KT2440R PP4392:: ΩKm, Kmr) | This study |
| E. coli BL21 (DE3) | F−, ompI, hsdSB (r−B m−B) | (65) |
| E. coli C41 (DE3) | F - ompT hsdSB (rB-mB-) gal dcm (DE3) | (66) |
| E.coli BL21 StarTM (DE3) | F- ompT hsdSB (rB-mB-) gal dcm rne131 (D3) | Invitrogen |
| E. coli DH5α | supE44 ΔlacU169 (ϕ80 lacZΔM15) hsdR1 recA1 endA1 gyrA96 thi1 relA1 | (67) |
| E. coli DH5α-TK1043 | DH5α with pGEM-T-3760, Apr | This study |
| E. coli DH5α-TK1044 | DH5α with pGEM-T-4392, Apr | This study |
| E. coli DH5α-TK1048 | DH5α with pChesi-3760, Kmr | This study |
| E. coli DH5α-TK1049 | DH5α with pChesi-4390, Kmr | This study |
| Plasmids | ||
| pET200/DTOPO | Kmr, protein expression vector | Invitrogen |
| pET28b(+) | Kmr, protein expression vector | Novagen |
| pET28b-WspC-Pp | Kmr, pET28b (+) derivative | (42) |
| pET28b-CheR2 | Kmr, pET28b (+) derivative | This work |
| pET28b-CheR3 | Kmr, pET28b (+) derivative | This work |
| pET200/D-TOPO | Kmr, protein expression vector | Invitrogen |
| pET200/D-TOPO-mcpT | mcpT gene inserted in pET200/D-TOPO, KmR | This work |
| pET28b-mcpS | mcpS gene inserted in pET28b (+), KmR | This work |
| pGRT1 | large self-transmissible plasmid present in P. putida DOT-T1E | (68) |
| pGEM-T | Cloning vector with polyA, ampr | Promega |
| pGEM-T-3760 | pGEM-T carrying between EcoRI and BamHI sites, a 400-bp chromosomal fragment from the P. putida KT2440R cheR3 gene obtained by PCR as an EcoRI-BamHI fragment, Apr | This study |
| pGEM-T-4392 | pGEM-T carrying between EcoRI and BamHI sites, a 400-bp chromosomal fragment from the P. putida KT2440R cheR2 gene obtained by PCR as an EcoRI-BamHI fragment, Apr | This study |
| pCHESIΩKm | Apr Kmr, pUNϕ18 with the HindIII insert from pHP45ϕKm (Ω-Km interposon) at the HindIII site, oriT RP4 | (69) |
| pChesi-3760 | pCHESIΩKm carrying between EcoRI and BamHI sites, a 400-bp chromosomal fragment from the P. putida KT2440R cheR3 gene obtained by PCR as an EcoRI-BamHI fragment | This study |
| pChesi-4392 | pCHESIΩKm carrying between EcoRI and BamHI sites, a 400-bp chromosomal fragment from the P. putida KT2440R cheR2 gene obtained by PCR as an EcoRI-BamHI fragment | This study |