Figure 5.

TrfA is active in the replication of the RK2 origin in crude extracts prepared from various E. coli clp mutants. The in vitro replication reactions with various amounts of wild-type His6-TrfA or His6-TrfA 254D/267L were performed under standard replication conditions as described in Materials and Methods with the use of extracts prepared from E. coli isogenic bacterial strains: (A) SG20250, (B) SG22080 (clpX∷kan), (C) SG22093 (clpA∷kan), and (D) SG 22100 (clpB∷kan). Replication activity was measured after incubation for 60 min at 32°C.