Reconstitution of F factor DNA replication in vitro with purified proteins.
作者： S. Zzaman , Mayuresh M. Abhyankar , Deepak Bastia
关键词: Origin recognition complex 、 Minichromosome maintenance 、 Ter protein 、 Control of chromosome duplication 、 Molecular biology 、 Replication factor C 、 Pre-replication complex 、 DNA replication 、 DnaA 、 Biology
摘要: Jacob, Brenner, and Cuzin pioneered the development of F plasmid as a model system to study replication control, these investigations led "replicon model" (Jacob, F., S., Cuzin, F. (1964) Cold Spring Harbor Symp. Quant. Biol. 28, 329-348). To elucidate further mechanism initiation this its we have reconstituted in vitro with 21 purified host-encoded proteins plasmid-encoded initiator RepE. The was specifically initiated at ori (oriV) required both bacterial protein DnaA wild type dimeric RepE inactive catalyzing replication, whereas monomeric mutant form called RepE(*) (R118P) capable vigorous replication. topology mostly Cairns form, fork movement unidirectional from right left. dependent on HU protein, structurally functionally related DNA bending IHF could not efficiently substitute for HU. priming DnaG primase. Many characteristics closely mimicked those vivo We believe that should be very useful unraveling control.
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