Figure 2
Western blot analysis of C-His 6 -VanY n and N-His 6 -VanY n from S. venezuelae recombinant strains. (A): analysis of samples corresponding to cellular and extracellular fractions from cultures of S. venezuelae recombinant strains grown in YEME for 72 hours. From S. venezuelae pIJ86ΩN-His6-vanY n : extracellular fraction (lane 1), soluble cell-free fraction (lane 2), insoluble cell-free fraction (lane 3) and cell wall fraction (lane 4); from S. venezuelae pIJ86ΩC-His6-vanY n : extracellular fraction (lane 5), soluble cell-free fraction (lane 6), insoluble cell-free fraction (lane 7) and cell wall fraction (lane 8). A crude extract from S. venezuelae cells carrying the empty vector was loaded as a negative control (CTRL). In each lane, samples corresponding to 100 μL of cell culture were loaded. Std reference protein: C-His6-VanYn from E. coli (5 μg, 25 kDa). The corresponding SDS-PAGE is shown in the Additional file 1: Figure S1. (B): quantitative analysis of recombinant VanYn present in each fraction, performed using the Quantity One program (Bio-Rad Laboratories, Milan, Italy) and His6-VanYn as comparative standard. Total protein content in each fraction was determined by Biuret method (□, solid line).