Fig. 3

SDS-PAGE and HPLC separation of HisrMlat1 from Origami strain cells. a SDS-PAGE. Left lane shows the molecular weight markers (kDa). Lane 1: the protein extract from cytoplasm. Lane 2: the protein content in the cytoplasmic soluble fraction (an arrow indicates the relative molecular size of the expected HisrMlat1). Lanes 3 and 4: the proteins that were not bound to the Ni-NTA column. Lanes 5 and 6: the eluates from the Ni-NTA after 250 mM imidazole. The protein bands corresponding to HisrMlat1 are indicated by an arrow. b rpHPLC. The product obtained from the affinity column was directly loaded into the C₁₈ reverse column (approx. 500 μg of protein). The fraction labelled with an asterisk had the molecular mass expected for the recombinant HisrMlat1. This fraction was lethal to mice. c Mass spectrometry. HisrMlat1 from either M15 or Origami strain cells has the same molecular mass, namely 8,499.6. The main m/z ions are shown [945.8]⁺⁹, [851.08]⁺¹⁰, [773.67]⁺¹¹