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Table 1 The strains, plasmids, and oligonucleotides used in this study

From: Overexpression and simple purification of the Thermotoga maritima 6-phosphogluconate dehydrogenase in Escherichia coli and its application for NADPH regeneration

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'-ccagtctactcgag gtgaaatctcatattggtctcatcggtc-3' pET-ci-wt6pgdh
CI-wt-R 5'-ccagtctagtcgac cctatctctccttcctcccag-3'  
CI-co-F 5'-ccagtcta ctcgag ggctcttccatgaaatcccacattggcctgatc-3' pET-ci-co6pgdh
CI-co-R 5'-ccagtctaggatcc tcaagtcgagccaatctccccctcctccc-3'  
NH-F 5'-gaggagggggagattggctaacatcaccaccaccattaag-3' pET-co6pgdh
NH-R 5'-cttaatggtggtggtgatgttagccaatctccccctcctc-3'  
G6P-F 5'-ccagtctactcgag ggctcttcc atgaagtgcagtctgggattg-3' pET-ci-g6pdh
G6P-R 5'-ccagtctagtcgac agttttctccattttctacc-3'  
  1. * Underlined nucleotide sequences indicate restriction endonuclease sites.