Table 4 Bacterial strains and plasmids used in this study

E. coli DH5α

supE44, DlacU169 ( f80 lacZ DM15), 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

P_lacUV5 T7 RNA pol, lacI ^q

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

P_lacUV5 T7 RNA pol, lacI ^q

This study

Ps. sp. strain VLB120Δbkd ΔilvE C18 T7

Ps. sp. strain VLB120Δbkd C18 ΔilvE, encoding branched-chain amino acid aminotransferase

This study

Ps. sp. strain VLB120Δbkd ΔpycAB C18 T7

Ps. sp. strain VLB120Δbkd C18 ΔpycAB, encoding pyruvate decarboxylase

This study

Ps. sp. strain VLB120Δbkd ΔleuA C18 T7

Ps. sp. strain VLB120Δbkd C18 ΔleuA, encoding 2-isopropylmalate synthase

This study

Ps. sp. strain VLB120Δbkd ΔpanB C18 T7

Ps. sp. strain VLB120Δbkd C18 ΔpanB, encoding 2-ketoisovalerate hydroxymethyltransferase

This study

Ps. sp. strain VLB120ΔbkdΔilvE ΔpanB C18 T7

Ps. sp. strain VLB120Δbkd ΔilvE C18 ΔpanB, encoding 2-ketoisovalerate hydroxymethyltransferase

This study

Ps. sp. strain VLB120Δbkd ΔilvE ΔleuA C18 T7

Ps. sp. strain VLB120Δbkd ΔilvE C18 ΔleuA, encoding 2-isopropylmalate synthase

This study

Ps. sp. strain VLB120Δbkd ΔilvE ΔleuA ΔpanB C18 T7

Ps. sp. strain VLB120Δbkd ΔilvE ΔleuA C18 T7ΔpanB, encoding 2-ketoisovalerate hydroxymethyltransferase

This study

Plasmids

Relevant characteristic(s)

Reference

pMA-RQ-kivd

Amp^r , Geneart delivery vector containing kivd

This study/Geneart

pCOM10

Km^r , 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

Cm^r, 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

Cm^r, Km^r, pBR22b-alsS with BoxI blunt end inserted Km^r

This study

pPAPC-km

pBR22b-alsS-km additionally containing ilvC

This study

pBR22b-eGFP

pCOM10 containing eGFP

This study

pBR22b-eGFP- km

Cm^r, Km^r pBR22b-eGFP with BoxI blunt end inserted Km^r

This study

pCOM8

Gm^r, 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

Gm^r, suicide vector, P15A ori sac B RP4 pBluescriptSK MCS

[68]

PJQ200SK-bkd-KO

Knock-out vector

This study

pJQhdp::km

Km^r, Gm^r, pJQ200SK with hdp disrupted by Km^r 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, P_lacUV5 T7 RNA pol, lacI ^q

[52]

pUC18 mini-Tn7-Gm-T7

pUC18, mini-Tn7-Gm, P_lacUV5 T7 RNA pol, lacI ^q

[52]

pTNS1

Amp^r, TnsABCD

[69]

pEMG

Km^r, oriR6K, lacZa with two flanking I-SceI sites

[64]

pSW-2

Gm^r, ori RK2, 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