Table 4.
Bacterial strains and plasmids used in this study
| Strains | Relevant characteristic(s) | Reference |
|---|---|---|
|
E. coli DH5α |
supE44, DlacU169 (f80 lacZDM15), hsdR17 (rk-mk+), recA1, endA1, thi1, gyrA, relA |
[63] |
|
E. coli DH5α ʎ-pir |
ʎ-pir lysogen of DH5a |
[64] |
|
Ps. sp. strain VLB120 |
Wildtype strain |
[65] |
|
Ps. sp. strain VLB120 T7 |
PlacUV5 T7 RNA pol, lacIq |
This study |
|
Ps. sp. strain VLB120Δbkd |
VLB120 with deletion of bkd, encoding subunit A of the branched-chain α-keto acid dehydrogenase |
This study |
|
Ps. sp. strain VLB120Δbkd A10, A21, B57, B83, C18, D67, D76, E82 |
NTG-mutants of Pseudomonas sp. strain VLB120 Δbkd |
This study |
|
Ps. sp. strain VLB120Δbkd C18 T7 |
PlacUV5 T7 RNA pol, lacIq |
This study |
|
Ps. sp. strain VLB120ΔbkdΔilvE C18 T7 |
Ps. sp. strain VLB120Δbkd C18 ΔilvE, encoding branched-chain amino acid aminotransferase |
This study |
|
Ps. sp. strain VLB120ΔbkdΔpycAB C18 T7 |
Ps. sp. strain VLB120Δbkd C18 ΔpycAB, encoding pyruvate decarboxylase |
This study |
|
Ps. sp. strain VLB120ΔbkdΔleuA C18 T7 |
Ps. sp. strain VLB120Δbkd C18 ΔleuA, encoding 2-isopropylmalate synthase |
This study |
|
Ps. sp. strain VLB120ΔbkdΔpanB C18 T7 |
Ps. sp. strain VLB120Δbkd C18 ΔpanB, encoding 2-ketoisovalerate hydroxymethyltransferase |
This study |
|
Ps. sp. strain VLB120ΔbkdΔilvEΔpanB C18 T7 |
Ps. sp. strain VLB120ΔbkdΔilvE C18 ΔpanB, encoding 2-ketoisovalerate hydroxymethyltransferase |
This study |
|
Ps. sp. strain VLB120ΔbkdΔilvEΔleuA C18 T7 |
Ps. sp. strain VLB120ΔbkdΔilvE C18 ΔleuA, encoding 2-isopropylmalate synthase |
This study |
|
Ps. sp. strain VLB120ΔbkdΔilvEΔleuAΔpanB C18 T7 |
Ps. sp. strain VLB120ΔbkdΔilvEΔleuA C18 T7ΔpanB, encoding 2-ketoisovalerate hydroxymethyltransferase |
This study |
|
Plasmids |
Relevant characteristic(s) |
Reference |
| pMA-RQ-kivd |
Ampr, Geneart delivery vector containing kivd |
This study/Geneart |
| pCOM10 |
Kmr , broad host range expression vector, alk promoter, oriT, alkS regulator gene, |
[66] |
| pCOM10-kivd |
pCOM10 containing kivd |
This study |
| pCOM10-eGFP |
pCOM10 containing eGFP |
This study |
| pBR22b |
Cmr, mob lacI
q
Km
r
PT7Ф10 |
[67] |
| pBR22b-kivd |
pBR22b containing kivd |
This study |
| pBR22b-alsS |
pBR22b containing alsS |
This study |
| pBR22b-ilvC |
pBR22b containing ilvC |
This study |
| pBR22b-ilvD |
pBR22b containing ilvD |
This study |
| pBR22b-alsS-km |
Cmr, Kmr, pBR22b-alsS with BoxI blunt end inserted Kmr |
This study |
| pPAPC-km |
pBR22b-alsS-km additionally containing ilvC |
This study |
| pBR22b-eGFP |
pCOM10 containing eGFP |
This study |
| pBR22b-eGFP-km |
Cmr, Kmr pBR22b-eGFP with BoxI blunt end inserted Kmr |
This study |
| pCOM8 |
Gmr, broad-host-range expression vector, alk promoter, oriT, alkS regulator gene |
[66] |
| pCOM8-T7-ilvD |
pCOM8 containing ilvD |
This study |
| pPDPK |
pCOM8-T7-ilvD containing kivd |
This study |
| pJQ200SK |
Gmr, suicide vector, P15A ori sacB RP4 pBluescriptSK MCS |
[68] |
| PJQ200SK-bkd-KO |
Knock-out vector |
This study |
| pJQhdp::km |
Kmr, Gmr, pJQ200SK with hdp disrupted by Kmr gene, flanked by loxP recombination sites |
Nick Wierckx (unpublished data) |
| pJTN-cre |
Gm
r
, encodes cre recombinase |
Nick Wierckx (unpublished data) |
| pUC18 mini-Tn7-Gm-T7 |
pUC18, mini-Tn7-Gm, PlacUV5 T7 RNA pol, lacIq |
[52] |
| pUC18 mini-Tn7-Gm-T7 |
pUC18, mini-Tn7-Gm, PlacUV5 T7 RNA pol, lacIq |
[52] |
| pTNS1 |
Ampr, TnsABCD |
[69] |
| pEMG |
Kmr, oriR6K, lacZa with two flanking I-SceI sites |
[64] |
| pSW-2 |
Gmr, oriRK2, xylS, Pm→ I-SceI |
[64] |
| pEMG-pycAB-KO |
Knock-out vector |
This study |
| pEMG-ilvE-KO |
Knock-out vector |
This study |
| pEMG-leuA-KO |
Knock-out vector |
This study |
| pEMG-panB-KO | Knock-out vector | This study |
Ampr: ampicillin resistance; Cmr: chloramphenicol resistance; Kmr: kanamycin resistance; Gmr: gentamicin resistance.