Table 1 E. coli strains, plasmids and oligonucleotides used
Name | Relevant Genotype | Reference |
|---|---|---|
Strains | Â | Â |
   DH10B | F- mcr A Δ(mrr-hsd RMS-mcr BC) ϕ80lac ZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ- rpsL nupG | Invitrogen |
   ElectroTen-Blue | Δ(mcrA)183 Δ(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac Kanr [F' proAB lacIqZ ΔM15 Tn10 (Tetr)] | Stratagene |
   MG1655 | F- λ- ilvG- rfb-50 rph-1 | ATCC 700926 |
   HCT 10 | MG1655 ΔendA ΔrecA (DE3) | This study |
   HCT 11 | MG1655 ΔendA ΔrecA ΔatoB (DE3) | This study |
   HCT 20 | MG1655 ΔendA ΔackA-pta ΔatoDA ΔpoxB (DE3) | This study |
   HCT 21 | MG1655 ΔendA ΔackA-pta ΔatoDA ΔpoxB ΔmetA Δtdh (DE3) | This study |
Plasmids | Â | Â |
   pETDuet-1 | ColE1(pBR322) ori, lacI, T7lac, AmpR | Novagen |
   pCDFDuet-1 | CloDF13 ori, lacI, T7lac, StrepR | Novagen |
   pCOLADuet-1 | COLA ori, lacI, T7lac, KanR | Novagen |
   pET-B | pETDuet-1 harboring bktB from R. eutropha H16 | This study |
   pET-PB-B a | pETDuet-1 harboring ptb-buk operon from C. acetobutylicum ATCC 824, and bktB from R. eutropha H16 | This study |
   pCDF-H | pCDFDuet-1 harboring hbd from C. acetobutylicum ATCC 824 | This study |
   pCDF-T-H a | pCDFDuet-1 harboring tesB from E. coli MG1655, and hbd from C. acetobutylicum ATCC 824 | This study |
   pCDF-P | pCDFDuet-1 harboring phaB from R. eutropha H16 | This study |
   pCDF-T-P a | pCDFDuet-1 harboring tesB from E. coli MG1655, and phaB from R. eutropha H16 | This study |
   pCOLA-Iec | pCOLADuet-1 harboring ilvA from E. coli MG1655 | This study |
   pCOLA-Icg | pCOLADuet-1 harboring ilvA from C. glutamicum | This study |
   pCOLA-Tec-Icg a | pCOLADuet-1 harboring thrABC operon from E. coli MG1655, and ilvA from C. glutamicum ATCC 13032 | This study |
   pCOLA-Tecm-Icg a | pCOLADuet-1 harboring thrAG1297ABC operon from E. coli ATCC 21277, and ilvA from C. glutamicum ATCC 13032 | This study |
Primers b | Sequence 5'→3' c |  |
   bktB_US_EcoRI | GAATTC ATGACGCGTGAAGTGGTAGTG | Sigma-Genosys |
   bktB_DS_XhoI | CTCGAG CGCAAGGCTAACCTCAGAT | Sigma-Genosys |
   hbd_US_NdeI | ATTCATATG AAAAAGGTATGTGTTATAGG | Sigma-Genosys |
   hbd_DS_AvrII | ATTCCTAGG CAGGTCGACTCTAGAACTTA | Sigma-Genosys |
   phaB_US_MfeI | ATTCAATTG ACGAAGCCAATCAAGGAG | Sigma-Genosys |
   phaB_DS_AvrII | ATTCCTAGG GGTCAGCCCATATGCAG | Sigma-Genosys |
   tesB_US_NcoI | ATTCCATGG GCATGAGTCAGGCGCTAA | Sigma-Genosys |
   tesB_DS_NotI | ATTGCGGCCGCG ACTCTAGAGACTTAATTGTG | Sigma-Genosys |
   ilvAec_US_NdeI | ATTACATATG GCTGACTCGCAAC | Sigma-Genosys |
   ilvAec_DS_AvrII | ATTACCTAGG CATTTTTCCCTAACC | Sigma-Genosys |
   ilvAcg_US_NdeI | ATTACATATG AGTGAAACATACGTGTC | Sigma-Genosys |
   ilvAcg_DS_AvrII | ATTACCTAGG CCTTCAGCTATGTTTA | Sigma-Genosys |
   thrABC_US_BamHI | ATTAGGATCC AAGGAGATATATCATGCGAGTGTTGAAG | Sigma-Genosys |
   thrABC_US_NcoI | ATTACCATGG GCATGCGAGTGTTGAAG | Sigma-Genosys |
   thrABC_DS_SalI | ATTAGTCGAC GATAATGAATAGATTTTACTGATG | Sigma-Genosys |