TABLE 4.
List of plasmids and strains used in the current study
| Strain or plasmid | Features or genotypea | Reference |
|---|---|---|
| Bacterial strains | ||
| E. coli DH5α λpir | Strain used for preparation and conjugative delivery of pSEVA212S-derived plasmids; F− ϕdlacZΔM15 ΔlacZYA-argF U169 deoR supE44 hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | Dunn et al. (78) |
| E. coli HB101 | Helper strain carrying pRK600 plasmid, used in triparental conjugation; F− thi-1 hsdS20 (rB–, mB–) supE44 recA13 ara-14 leuB6 proA2 lacY1 galK2 rpsL20 (Strr) xyl-5 mtl-1 | Boyer and Roulland-Dussoix (79) |
| E. coli BL21(DE3) | Strain used for high-level T7 expression of cloned oxidoreductases; fhuA2 [lon] ompT gal (λDE3) [dcm] ΔhsdS λ sBamHIo ΔEcoRI-B int::(lacI::PlacUV5::T7 gene1) i21 Δnin5 | Studier and Moffatt (71) |
| Pseudomonas putida KT2440 | Model organism | DSM 6125 |
| P. putida KT2440 ΔPP_2426 | Deletion mutant without calA (PP_2426) ORF | This study |
| P. putida KT2440 ΔPP_3839 | Deletion mutant without PP_3839 ORF | This study |
| P. putida GN442 | Engineered strain for production of vanillin from ferulate; Δupp ΔPP_0166-0168 Δvdh ΔPP_3827-3832 ΔPP_2680 ΔPP_0545 ΔPP_1948 lacIq-Ptac-ech-fcs | Graf and Altenbuchner (26) |
| P. putida GN442 ΔPP_2426 | Optimized strain for high-yield production of vanillin from ferulate; Δupp ΔPP_0166-0168 Δvdh ΔPP_3827-3832 ΔPP_2680 ΔPP_0545 ΔPP_1948 lacIq-Ptac-ech-fcs ΔPP_2426 | This study |
| Pseudomonas sp. 9.1 | Environmental isolate | Ravi et al. (19), DSM 105530 |
| Plasmids | ||
| pNIC28-Bsa4 | Plasmid for ligation-independent cloning and T7 expression of cloned genes, Kmr | Savitsky et al. (80) |
| pNIC28-Bsa4KpnI | KpnI-digested and religated version of pNIC28-Bsa4 used as negative-control plasmid, Kmr | This study |
| pNIC28-Bsa4-calA | Clone of PP_2426 in pNIC28-Bsa4, Kmr | This study |
| pNIC28-Bsa4-PP_3839 | Clone of PP_3839 in pNIC28-Bsa4, Kmr | This study |
| pNIC28-Bsa4-FEZ21_09870 | Clone of FEZ21_09870 in pNIC28-Bsa4, Kmr | This study |
| pSEVA212S | Suicide plasmid for integration in Pseudomonas genomes, Kmr | Martínez-García and de Lorenzo (75) |
| pSEVA212S-ΔcalA | pSEVA212S containing the upstream and downstream regions of calA assembled together, Kmr | This study |
| pSEVA212S-ΔPP_3839 | pSEVA212S containing the upstream and downstream regions of PP_3839 assembled together, Kmr | This study |
| pRK600 | Helper plasmid used in triparental mating between E, coli and Pseudomonas strains, Cmr | Finan et al. (81) |
| pSEVA128S | Plasmid carrying the gene encoding I-SceI endonuclease, Apr | Martínez-García and de Lorenzo (75) |
a
Strr, streptomycin resistance; Kmr, kanamycin resistance; Cmr, chloramphenicol resistance; Apr, apramycin resistance.