Table 1 Bacterial strains and plasmids used in this study
Strain or plasmid | Relevant characteristics | Source or Reference |
|---|---|---|
E. coli | ||
DH5α | F− Φ80dlac∆(lacZ)M15 ∆(lacZYA-argF) U169 endA1 recA1 hsdR17 (rK−, mK+) deoR thi-1 phoA supE44 λ−gyrA96 relA1; strain used for cloning procedures | [26] |
C. glutamicum | ||
ATCC13032 (WT) | Biotin-auxotrophic wild type | [78] |
WT ΔTRP ΔtrpP (WT* ΔTRP in the original publication) | WT with an in-frame deletion of ΔtrpP (cg3357) ΔtrpE (cg3359) ΔtrpG (cg3360) ΔtrpD (cg3361) ΔtrpCF (cg3362) ΔtrpB (cg3363) ΔtrpA (cg3364) | [13] |
C1* ΔTRP ΔtrpP (C1* ΔTRP in the original publication) | C1* with an in-frame deletion of ΔtrpP (cg3357) ΔtrpE (cg3359) ΔtrpG (cg3360) ΔtrpD (cg3361) ΔtrpCF (cg3362) ΔtrpB (cg3363) ΔtrpA (cg3364) | [13] |
C1* ΔTRP ΔtrpP evo1 | Evolved C1* ΔTRP ΔtrpP strain number 1 with mutation SufRL25P | This study |
C1* ΔTRP ΔtrpP evo2 | Evolved C1* ΔTRP ΔtrpP strain number 2 with mutation SufRR77G | This study |
C1* ΔTRP ΔtrpP evo3 | Evolved C1* ΔTRP ΔtrpP strain number 3 with mutation SufRQ193* | This study |
WT ΔtrpP | WT with an in-frame deletion of ΔtrpP (cg3357) | This study |
WT ΔTRP | WT with an in-frame deletion of ΔtrpE (cg3359) ΔtrpG (cg3360) ΔtrpD (cg3361) ΔtrpCF (cg3362) ΔtrpB (cg3363) ΔtrpA (cg3364) | This study |
WT ΔTRP ΔtrpP ΔsufR | WT ΔTRP ΔtrpP with a partial deletion of sufR (cg1765) until the sufB (cg1764) promoter site with an inserted stop codon at the end of sufR | This study |
WT ΔTRP ΔtrpP SufRL25P | WT ΔTRP ΔtrpP with point mutation L25P in SufR | This study |
WT ΔTRP ΔtrpP SufRQ193* | WT ΔTRP ΔtrpP with point mutation Q193* in SufR | This study |
WT ΔTRP ΔtrpP ΔPsufB::Ptuf | WT ΔTRP ΔtrpP with an exchange of sufR (cg1765) and the native sufB promoter sequence with the tuf promoter (cg0587) | This study |
WT ΔTRP ΔtrpP ΔPsufB::PdapA | WT ΔTRP ΔtrpP with an in frame exchange of sufR (cg1765) including the native sufB promoter sequence with the dapA promoter (cg2161) | This study |
WT ΔtrpP ΔsufR | WT ΔtrpP with an partial deletion of sufR (cg1765) until the sufB (cg1764) promoter site with an inserted stop codon at the end of sufR | This study |
WT TRP+ | WT with point mutations trpLfbr (with mutation of the third of three tandem Trp codons of trpL TGG ➜ TGA) and TrpES38R (Cg3359) | This study |
WT TRP++ | WT TRP+with point mutation TrpDA162E (Cg3361) | This study |
WT TRP+++ | WT TRP++ with in-frame deletion of aroP (cg1257) | This study |
WT TRP+++ ΔsufR | WT TRP+++ with a partial deletion of sufR (cg1765) until the sufB (cg1764) promoter site with an inserted stop codon at the end of sufR | This study |
WT TRP+++ ΔtrpP | WT TRP+++ with an in-frame deletion of trpP (cg3357) | This study |
WT TRP+++ ΔtrpP ΔsufR | WT TRP+++ ΔtrpP with an partial deletion of sufR (cg1765) until the sufB (cg1764) promoter site with an inserted stop codon at the end of sufR | This study |
Plasmids | ||
pK19mobsacB | KanR.; plasmid for allelic exchange in C. glutamicum; (pK18 oriVE.c., sacB, lacZα) | [27] |
pK19mobsacB-TrpLfbr TrpES38R | KanR.; pK19mobsacB derivative for mutation of trpL (TGGâ—ŠTGA) and TrpE (Cg3359) S38R in C. glutamicum | [13] |
pK19mobsacB-ΔTRPv2_(trpP) | KanR.; pK19mobsacB derivative for in-frame deletion of TRP-operon (cg3359-cg3364) without disrupting trpP (cg3357) in C. glutamicum | This study |
pK19mobsacB-ΔtrpPv2 (TrpLfbr TrpES38R) | KanR.; pK19mobsacB derivative for in-frame deletion of trpP (cg3357) in C. glutamicum WT TRP+without disrupting the inserted mutations in trpL and TrpE (Cg3359) | This study |
pK19mobsacB-ΔtrpP | KanR.; pK19mobsacB derivative for in-frame deletion of trpP (cg3357) in C. glutamicum | This study |
pK19mobsacB-ΔaroP | KanR.; pK19mobsacB derivative for in-frame deletion of aroP (cg1257) in C. glutamicum | This study |
pK19mobsacB-ΔsufR | KanR.; pK19mobsacB derivative for partial in-frame deletion of sufR (cg1765) with an inserted stop codon at the end of sufR in C. glutamicum | This study |
pK19mobsacB-SufRL25P | KanR.; pK19mobsacB derivative for mutation of SufR (Cg1765) L25P in C. glutamicum | This study |
pK19mobsacB-SufRQ193* | KanR.; pK19mobsacB derivative mutation of SufR (Cg1765) Q193* in C. glutamicum | This study |
pK19mobsacB-TrpDA162E | KanR.; pK19mobsacB derivative for mutation of TrpD (Cg3361) A162E in C. glutamicum | This study |
pK19mobsacB-ΔsufRΔPsufB::Ptuf | KanR.; pK19mobsacB derivative for deletion of sufR (cg1765) with integration of Ptuf (cg0587) in front of the SUF-cluster in C. glutamicum | This study |
pK19mobsacB-ΔsufRΔPsufB::PdapA | KanR.; pK19mobsacB derivative for deletion of sufR (cg1765) with integration of Ptuf (cg2161) in front of the SUF-cluster in C. glutamicum | This study |
pPREx2 | KanR; C. glutamicum/E. coli shuttle vector for regulated gene expression using the Ptac promoter | [24] |
pPREx2-trpP | KanR; pPREx2 derivative for trpP (cg3357) expression under control of Ptac | This study |
pJC1 | KanR; E. coli/C. glutamicum shuttle (oriVE.coli, oriVC.glutamicum) | [28] |
pJC1-Venus-Term | KanR; pJC1 derivative carrying the venus coding sequence and additional terminators, used as PCR template for venus. | [79] |
pJC1-PsufR-venus | KanR; pJC1 derivative with venus fluorescent protein under control of the native sufR (cg1765) promoter | This study |
pJC1-PsufR-venus-MutTSS1 | KanR; pJC1 derivative with venus fluorescent protein under control of the sufR (cg1765) promoter with mutated − 10 region of TSS1 (AAC ◊ CTG) | This study |
pJC1-PsufR-venus-MutTSS2 | KanR; pJC1 derivative with venus fluorescent protein under control of the sufR (cg1765) promoter with mutated − 10 region of TSS2 (GTG ◊ TAC) | This study |