RecO at 150 μg and RecR at 150 μg, which
correspond to 56 μM and 68 μM, respectively, or RecF at 150 μg,
RecO at 150 μg, and RecR at 150 μg, which correspond to 38 μM, 56
μM, and 68 μM, respectively, were mixed in buffer I, and incubated
on ice for 30 min prior to applying on Ssb affinity column. Bound
proteins were eluted in linear gradient of salt. (A)
RecO–RecR complex on Ssb affinity column. (B)
RecF–RecO–RecR complex on Ssb affinity column. (C) Peak
fractions were analyzed by SDS/PAGE and proteins were visualized by
Coomassie blue staining. (A) Peak fractions obtained
when RecO–RecR was applied to the Ssb column. (B) The peak
fraction obtained when the RecF–RecO–RecR complex was applied. Each
lane represent proteins present in corresponding peak fractions.