Figure 5.
The THI4 homolog (HvThi4) and its C165A variant are synthesized with C-terminal StrepII tags as stable derivatives in Haloferax volcanii . Production of the HvThi4-StrepII and HvThi4 C165A-StrepII proteins in the trans complemented strains (NC1011-pJAM2821 and -pJAM2822, respectively) was detected by anti-StrepII Western blotting (0.065 OD600 units of cells per lane). Equivalent protein loading was assessed by staining blots with Ponceau Red S (not shown) and parallel SDS-PAGE gels with Coomassie Blue R-250 (CB stain, 37–50 kDa range of gel shown).