Table 1.
The strains, plasmids, and oligonucleotides used in this study
| Description | Contents | Reference/sources |
| Strain | ||
| E. coli Bl21star(DE3) | B F- ompT hsdSB(rB-mB-) gal dcm rne131 (DE3) | Invitrogen |
| E. coli Bl21(DE3)-RIL | B F- ompT hsdS(rB-mB-) dcm Tetr gal λ(DE3) endA Hte [argU ileY leuW Camr] |
Stratagene |
| Plasmid | ||
| pCIP | Ampr, T7 promoter, lacO, ColE1 ori, parental DNA, replacing pgm gene with wt6pgdh, co6pgdh or g6pdh |
[44] |
| pET21a | Epoch Biolabs | |
| pET-trx-wt6pgdh | Ampr, T7 promoter, lacO, ColE1 ori, Trx-wt6pgdh | This study |
| pET-ci-wt6pgdh | wt6pgdh gene subcloned into pCIP | This study |
| pET-ci-co6pgdh | co6pgdh gene subcloned into pCIP | This study |
| pET-co6pgdh-his | co6pgdh gene with C-terminal (His)6 cloned into pET21a | |
| pET-co6pgdh | Removed C-terminal (His)6 from pET-co6pgdh-his | This study |
| pET-ci-g6pdhp | Expression of T. maritima g6pdh | This study |
| pET26b-xr | Expression of N. crassa xylose reductase | [57] |
| Primers* | Final plasmid | |
| Trx-F | 5'-caccatggtgaaatctcatattggtctcatcggtc-3' | pET-trx-wt6pgdh |
| Trx-R | 5'-tcatcctatctctccttcctcccagttg-3' | |
| CI-wt-F | 5'-ccagtctactcgaggtgaaatctcatattggtctcatcggtc-3' | pET-ci-wt6pgdh |
| CI-wt-R | 5'-ccagtctagtcgaccctatctctccttcctcccag-3' | |
| CI-co-F | 5'-ccagtcta ctcgagggctcttccatgaaatcccacattggcctgatc-3' | pET-ci-co6pgdh |
| CI-co-R | 5'-ccagtctaggatcctcaagtcgagccaatctccccctcctccc-3' | |
| NH-F | 5'-gaggagggggagattggctaacatcaccaccaccattaag-3' | pET-co6pgdh |
| NH-R | 5'-cttaatggtggtggtgatgttagccaatctccccctcctc-3' | |
| G6P-F | 5'-ccagtctactcgagggctcttcc atgaagtgcagtctgggattg-3' | pET-ci-g6pdh |
| G6P-R | 5'-ccagtctagtcgacagttttctccattttctacc-3' |
* Underlined nucleotide sequences indicate restriction endonuclease sites.