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Point mutations altering the Rcl1p–Bms1p interaction in vitro. Point mutations have been introduced in Rcl1p in order to affect protein–protein interactions. Wild-type and mutant Rcl1p were co-expressed in E. coli with N-terminally His-fused Bms1p and pull-down experiments were performed with cobalt-affinity resin. The presence of the proteins was analysed by SDS-PAGE and Coomassie-blue staining. The amount of Rcl1p in crude extracts was controlled using anti-Rcl1p antibodies.
