Table 2 Plasmids and strains used in this study.
Plasmid or strain | Relevant characteristics | Source |
|---|---|---|
Plasmid | ||
pEASY-Blunt | f1 ori, T7 promoter, KanR and AmpR | Transgen |
pETDuet-1 | f1 ori, T7 promoter, double T7 promoter, AmpR | Novagen |
pACYCDuet-1 | p15A ori, double T7 promoter, CmR | Novagen |
tyrAfbr-aroGfbr-tktA-ppsA/pCOLA | pCOLADuet-1 contained the tyrAfbr, aroGfbr, tktA and ppsA genes | [25] |
tal/pACYC | pACYCDuet-1 contained the tal gene | This study |
sam8/pACYC | pACYCDuet-1 contained the sam8 gene | This study |
tal-hpaBC/pACYC | pACYCDuet-1 contained the tal and hpaBC genes | This study |
sam8-sam5/pACYC | pACYCDuet-1 contained the sam8 and sam5 genes | This study |
tal-hpaBC-comt/pACYC | pACYCDuet-1 contained the tal, hpaBC and comt genes | This study |
sam8-sam5-comt/pACYC | pACYCDuet-1 contained the sam8, sam5 and comt genes | This study |
fcs-ech/pET | pETDuet-1 contained the fcs and ech genes | This study |
fcs-ech-sam5/pET | pETDuet-1 contained the fcs, ech and sam5genes | This study |
Strain | ||
E. coli DH5a | Cloning host | Invitrogen |
E. coli BL21(DE3) | Expression host | Novagen |
S. espanaensis DSM 44229 | Used for cloning the sam8 and sam5 genes | From DSMZ |
R. sphaeroides DSM 158 | Used for cloning the tal gene | From DSMZ |
Tyrosine overproducing strain | ΔtyrR harboring pCOLA-TATP | [25] |
VT-1 | E. coli BL21(DE3) harboring sam8-sam5-comt/pACYC | This study |
VT-2 | E. coli BL21(DE3) harboring tal-hpaBC-comt/pACYC | This study |
VT-3 | E. coli BL21(DE3) harboring fcs-ech/pET | This study |
VT-4 | E. coli BL21(DE3) harboring sam8-sam5-comt/pACYC and fcs-ech/pET | This study |
VT-5 | E. coli BL21(DE3) harboring sam8-sam5-comt/pACYC and fcs-ech-sam5/pET | This study |
VT-6 | Tyrosine overproducing strain harboring sam8-sam5-comt/pACYC and fcs-ech/pET | This study |
