Biomphalysin, a New β Pore-forming Toxin Involved in Biomphalaria glabrata Immune Defense against Schistosoma mansoni
Figure 5
Analysis of rBiomphalysin expressed in vitro using wheat germ extracts (RTS 500).
A. The amount of rBiomphalysin production was assessed by comparing the RTS 500 His-tagged Biomphalysin reaction (lanes 2 and 4) with 1 and 5 µl of RTS 500 control reactions (lanes 1 and 3) in Coomassie blue-stained, 12% SDS-PAGE gels. Values on the right indicate the masses of the molecular weight markers, whereas the arrow indicates the position of the His-tagged Biomphalysin band (lanes 2 and 4). B. Western blot analysis of RTS 500 control (lane 1) and Biomphalysin (lane 2) reactions. Five microliters of each reaction were separated by SDS-PAGE on a 12% gel and then electrotransferred onto a nitrocellulose membrane. The membrane was then incubated sequentially with a monoclonal anti-His6 antibody and a goat anti-mouse IgG, and immunoreactive proteins were visualized by ECL.