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Nucleic Acids Research, 1982, Vol. 10, No. 15 4687-4702
© 1982

MOLECULAR BIOLOGY

Replication of the broad host range plasmid RSF1010 in cell-free extracts of Escherichia coli and Pseudomonas aeruginosa

Ramon Diaz* and Walter L. Staudenbauer+

Max-Planck-Institut für Molekulare Genetik Abt. Schuster, Ihnestrasse 63-73, D-1000 Berlin 33, FRG

Received June 8, 1982. Revised July 6, 1982. Accepted July 6, 1982.

Replication of exogenous RSF1010 DNA can be carried out by soluble enzyme systems from Escherichia coli and Pseudomonas aeruginosa. It requires the function of RSF1010-encoded replication protein(s), which is not expressed in extracts from plasmid-free bacteria. In contrast to previously described in vitro systems for plasmid replication, initiation of RSF1010 DNA synthesis is independent of transcription catalyzed by host RNA polymerase. This is indicated by the insensitivity of RSF1010 replication to rifampicin as well as to RNA polymerase antibodies. It is proposed that a host RNA polymerase transcription-independent initiation mechanism might be a general property of broad host range plasmids.

*Present address: Centro de Investigaciones Biologicas (C.S.I.C.), Instituto de Biologia Celular, C/Velazquez 144, Madrid-6, Spain.

+Present address: Lehrstuhl für Mikrobiologie, Technische Universität München, Arcisstrasse 21, D-8000 München 2, FRG.


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