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- W2072687774 abstract "To elucidate the replication mechanism of a ColE1-type plasmid in RNase H-deficient (rnh−) strains of Escherichia coli, we constructed plasmid derivatives that deleted the whole, or a part, of the 5′-AAAAA-3′ sequence (positions −3 to +2) that acts as the origin of replication in vivo and in vitro in the presence of RNase H. The activity of plasmid replication in rnh+ cells was found to be reduced by alterations of the AAAAA sequence. The activity could be restored when the derivatives, retaining the upstream sequence down to −8, regained a sequence containing at least two A residues in the region from −3 to +2. By contrast, replication in rnh+ cells was maintained at high levels even when the deletion included the AAAAA sequence and extended up to position −7. The activity in rnh− cells decreased as deletions proceeded to −8 and further up to −17, and was abolished completely by further upward deletions. We concluded that in rnh− cells the plasmid replicates by a mechanism that operates only when RNase H is inactive. This RNase H-sensitive replication in rnh− cells seems to require the RNA-DNA hybrid formation that is also required for RNase H-dependent replication in rnh+ cells. The hybrid formation probably contributes by unwinding a portion of DNA from which replication can be initiated." @default.
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- W2072687774 date "1987-11-01" @default.
- W2072687774 modified "2023-10-10" @default.
- W2072687774 title "Nucleotide sequences required for a ColE1-type plasmid to replicate in Escherichia coli cells with or without RNase H" @default.
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- W2072687774 doi "https://doi.org/10.1016/0022-2836(87)90308-1" @default.
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