Table 3.
Strains, plasmids, and primers used in this study.
| Strain E. coli | Relevant Characteristic(s) or Sequence | Source and/or Reference |
|---|---|---|
| BW25113 | F−, DE(araD-araB)567, lacZ4787(del)::rrnB-3, LAM−, rph-1, DE(rhaD-rhaB)568, hsdR514 | [34] |
| BL21(DE3)Δzwf::kanr | F− ompT gal dcm lon hsdSB(rB− mB−) λ(DE3 (lacI lacUV5-T7 gene 1 ind1 sam7 nin5)) Δzwf-777::kan | [23] |
| Plasmids | ||
| pETg6pd | pET-3a carrying the human g6pd gene, AmpR | [23] |
| pETgR257L | pET-3a carrying the human g6pd gene with a R257L mutation in the G6PD protein, AmpR | This study |
| pETgL128P | pET-3a carrying the human g6pd gene with an L128P mutation in the G6PD protein, AmpR | This study |
| pETgV291M | pET-3a carrying the human g6pd gene with an V291M mutation in the G6PD protein, AmpR | This study |
| Mutagenesis | Primer Sequence | |
| R257L fw | 5′-GGATCATCCTGGACGTGATG-3′ | This study |
| R257L rev | 5′-CATCACGTCCAGGATGATTCC-3′ | This study |
| L128P fw | 5′-GAATGCCCTCCACCTGGG-3′ | This study |
| L128P rev | 5′-CTTACGGGAGGTGGACCC-3′ | This study |
| V291M fw | 5′-GAGAAGGTCAAGATGTTGAAATG-3′ | This study |
| V291M rev | 5′-CATTTCAACATCTTCACCTTCTC-3′ | This study |
The locations of the mutagenic oligonucleotides are in bold.