TABLE 1.
Bacterial strains and plasmids used in this study
| Strains and plasmids | Relevant genotype and characteristics | Reference |
|---|---|---|
| E. coli | ||
| JM105 | thi rpsL (Strr) endA sbcB15 sbcC hsdR4 (rK− mK−) Δ(lac-proAB) [F′ traD36 lacIq Δ(lacZ)M15 proA+B+] | 29 |
| HB101 | SmrrecA thi pro leu hsdRM+ | 14 |
| MV1190 | Δ(lac-proAB) Δ(srl-recA)306::Tn10 [F′ traD36 proAB lacIq Δ(lacZ)M15] thi supE | 27 |
| MV1190(λ-pir) | As MV1190, lysogenized with λ-pir phage | 11 |
| P. putida | ||
| KT2442 | hsdR Rifr | 11 |
| KT2442 (lacIq) | As KT2442 with a mini-Tn5-lacIq cassette inserted into the chromosome; Rifr | Unpublished |
| JB279 | KT2442 (lacIq) × HB101(RK600) × MV1190(λ-pir)(pJBA29)a; KT2442 (lacIq) with a mini-Tn5-Km-T1-T0-gfpmut3*-PA1/04/03 cassette from pJBA29 randomly inserted into the chromosome; Rifr Kmr | This study |
| JB167 | KT2442 (lacIq) × HB101(RK600) × MV1190(λ-pir)(pJBA115)a; KT2442 (lacIq) with a mini-Tn5-Km-T1-T0-gfp(LAA)-PA1/04/03 cassette from pJBA115 randomly inserted into the chromosome; Rifr Kmr | This study |
| JB391 | KT2442 (lacIq) × HB101(RK600) × MV1190(λ-pir)(pJBA117)a; KT2442 (lacIq) with a mini-Tn5-Km-T1-T0-gfp(LVA)-PA1/04/03 cassette from pJBA117 randomly inserted into the chromosome; Rifr Kmr | This study |
| JB340 | KT2442 (lacIq) × HB101(RK600) × MV1190(λ-pir)(pJBA119)a; KT2442 (lacIq) with a mini-Tn5-Km-T1-T0-gfp(AAV)-PA1/04/03 cassette from pJBA119 randomly inserted into the chromosome; Rifr Kmr | This study |
| JB396 | KT2442 (lacIq) × HB101(RK600) × MV1190(λ-pir)(pJBA121)a; KT2442 (lacIq) with a mini-Tn5-Km-T1-T0-gfp(ASV)-PA1/04/03-cassette from pJBA121 randomly inserted into the chromosome; Rifr Kmr | This study |
| Plasmids | ||
| pUC18Not | Apr; cloning vector | 11 |
| pQE70 | Apr; donor of RBSII-T0-T1 cassette | 21 |
| pUHE24-2 | Apr; donor of PA1/04/03 promoter | 17 |
| pJBA23 | Apr; pUC18Not-RBSII-T0-T1 | This study |
| pJBA24 | Apr; pUC18Not-PA1/04/03-RBSII-T0-T1 | This study |
| pJBA27 | Apr; pUC18Not-PA1/04/03-RBSII-gfpmut3*-T0-T1 | This study |
| pJBA110 | Apr; pUC18Not-PA1/04/03-RBSII-gfp(LAA)-T0-T1 | This study |
| pJBA111 | Apr; pUC18Not-PA1/04/03-RBSII-gfp(LVA)-T0-T1 | This study |
| pJBA112 | Apr; pUC18Not-PA1/04/03-RBSII-gfp(AAV)-T0-T1 | This study |
| pJBA113 | Apr; pUC18Not-PA1/04/03-RBSII-gfp(ASV)-T0-T1 | This study |
| pUT-miniTn5-Km | Apr Kmr; delivery plasmid for mini-Tn5-Km | 9 |
| pJBA28 | Apr; Kmr; delivery plasmid for mini-Tn5-Km-PA1/04/03-RBSII-gfpmut3*-T0-T1 | This study |
| pJBA29 | As pJBA28, but with the PA1/04/03-RBSII-gfpmut3*-T0-T1 cassette of pJBA27 in the opposite orientation | This study |
| pJBA114 | Apr Kmr; delivery plasmid for mini-Tn5-Km-PA1/04/03-RBSII-gfp(LAA)-T0-T1 | This study |
| pJBA115 | As pJBA114, but with the PA1/04/03-RBSII-gfp(LAA)-T0-T1 cassette of pJBA110 in the opposite orientation | This study |
| pJBA116 | Apr Kmr; delivery plasmid for mini-Tn5-Km-PA1/04/03-RBSII-gfp(LVA)-T0-T1 | This study |
| pJBA117 | As pJBA116, but with the PA1/04/03-RBSII-gfp(LVA)-T0-T1 cassette of pJBA111 in the opposite orientation | This study |
| pJBA118 | Apr Kmr; delivery plasmid for mini-Tn5-Km-PA1/04/03-RBSII-gfp(AAV)-T0-T1 | This study |
| pJBA119 | As pJBA118, but with the PA1/04/03-RBSII-gfp(AAV)-T0-T1 cassette of pJBA112 in the opposite orientation | This study |
| pJBA120 | Apr Kmr; delivery plasmid for mini-Tn5-Km-PA1/04/03-RBSII-gfp(ASV)-T0-T1 | This study |
| pJBA121 | As pJBA120, but with the PA1/04/03-RBSII-gfp(ASV)-T0-T1 cassette of pJBA113 in the opposite orientation | This study |
| RK600 | Cmrori ColE1 RK2-Mob+ RK2-Tra+; helper plasmid in triparental matings | 14 |
a
Three-factor matings were performed as described previously by Kristensen et al. (16).