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- W2470481570 abstract "The origin of replication, oriC, of the Esche- richia coli chromosome was mapped within a DNA segment of 422 base pairs. The nucleotide sequence of this segment was determined. The source of DNA for the sequence analysis was a minichromosome constructed in vivo, consisting exclusively of chromosomal DNA and a minichromosome constructed by cloning in vitro. The nucleotide sequence of the replication origin is characterized by a high degree of repetitiveness due to both inverted and direct repeats. Sequence homologies were found between portions of the replication origins of E. coli and phages X and G4. This suggests similarities in some steps in the initiation of replication of the different replicons. Replication of the circular chromosome of Escherichia coli is initiated at a fixed site, the origin of replication, for which the symbol oriC has been introduced (cf. refs. 1, 2). We define oriC here as the minimal segment of DNA that is able to promote E. coli specific initiation of replication. Isolation and charac- terization of specialized transducing phages Xasn carrying the origin of replication of the E. coli chromosome has led to the allocation of oriC between the genetic markers uncB and asn at 82.5 min on the circular genetic map (1, 2). This is in accor- dance with the localization of oriC obtained by other genetic techniques (3, 4). The replication origin can be placed within a segment of 1500 base pairs of DNA counterclockwise of the asn gene (1, 2). A 5.9-megadalton (9.0 kilobase pairs) EcoRI fragment from this region of the chromosome (ref. 5; Fig. 1A) has been cloned in minichromosomes, plasmids that contain oriC as the sole replication origin (3, 6). The position of oriC has been narrowed down further to a segment of 500 base pairs by means of deletions introduced into minichromosomes in vitro by various restriction endonucleases (6, 7). Here we present the nucleotide sequence of the DNA seg- ment containing oriC. As a substrate for the sequence analysis, we constructed by genetic techniques (in vivo) a minichro- mosome 2.65 megadaltons (4.0 kilobase pairs) in size. This consisted exclusively of chromosomal DNA that included, be- sides oriC, the asn gene. As an alternative source of DNA we used minichromosomes constructed independently by means of in vitro techniques. Results complementary to our data are described in an ac-" @default.
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- W2470481570 date "2016-01-01" @default.
- W2470481570 modified "2023-09-27" @default.
- W2470481570 title "(initiation of DNA replication/minichromosomes/phage X/phagi" @default.
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