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Replication of ColE2 and ColE3 plasmids: in vitro replication dependent on plasmid-coded proteins.
Mol Gen Genet. 1989 Oct; 219(1-2):249-55.MG

Abstract

We developed an in vitro replication system for ColE2 and ColE3 plasmids using cell extracts prepared from bacteria with or without these plasmids. DNA synthesis depended on host DNA polymerase I and was sensitive to rifampicin and chloramphenicol. Preincubation of the extracts with plasmid DNA, however, allowed replication of template DNA added subsequently in a plasmid-specific manner in the presence of rifampicin and chloramphenicol. The plasmid-specified trans-acting factor(s) was detected in cell extracts from bacteria carrying a recombinant plasmid with the region of ColE2 or ColE3 encoding the Rep protein. The plasmid-specified factor(s) consisted at least in part of protein, probably the Rep protein. In vitro replication started within a region of ColE2 or ColE3 containing the smallest cis-acting segment essential for in vivo replication and proceeded in a fixed direction.

Authors+Show Affiliations

Department of Biology, Faculty of Science, Osaka University, Japan.No affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

2693943

Citation

Itoh, T, and T Horii. "Replication of ColE2 and ColE3 Plasmids: in Vitro Replication Dependent On Plasmid-coded Proteins." Molecular & General Genetics : MGG, vol. 219, no. 1-2, 1989, pp. 249-55.
Itoh T, Horii T. Replication of ColE2 and ColE3 plasmids: in vitro replication dependent on plasmid-coded proteins. Mol Gen Genet. 1989;219(1-2):249-55.
Itoh, T., & Horii, T. (1989). Replication of ColE2 and ColE3 plasmids: in vitro replication dependent on plasmid-coded proteins. Molecular & General Genetics : MGG, 219(1-2), 249-55.
Itoh T, Horii T. Replication of ColE2 and ColE3 Plasmids: in Vitro Replication Dependent On Plasmid-coded Proteins. Mol Gen Genet. 1989;219(1-2):249-55. PubMed PMID: 2693943.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Replication of ColE2 and ColE3 plasmids: in vitro replication dependent on plasmid-coded proteins. AU - Itoh,T, AU - Horii,T, PY - 1989/10/1/pubmed PY - 1989/10/1/medline PY - 1989/10/1/entrez SP - 249 EP - 55 JF - Molecular & general genetics : MGG JO - Mol. Gen. Genet. VL - 219 IS - 1-2 N2 - We developed an in vitro replication system for ColE2 and ColE3 plasmids using cell extracts prepared from bacteria with or without these plasmids. DNA synthesis depended on host DNA polymerase I and was sensitive to rifampicin and chloramphenicol. Preincubation of the extracts with plasmid DNA, however, allowed replication of template DNA added subsequently in a plasmid-specific manner in the presence of rifampicin and chloramphenicol. The plasmid-specified trans-acting factor(s) was detected in cell extracts from bacteria carrying a recombinant plasmid with the region of ColE2 or ColE3 encoding the Rep protein. The plasmid-specified factor(s) consisted at least in part of protein, probably the Rep protein. In vitro replication started within a region of ColE2 or ColE3 containing the smallest cis-acting segment essential for in vivo replication and proceeded in a fixed direction. SN - 0026-8925 UR - https://www.unboundmedicine.com/medline/citation/2693943/Replication_of_ColE2_and_ColE3_plasmids:_in_vitro_replication_dependent_on_plasmid_coded_proteins_ L2 - https://doi.org/10.1007/BF00261184 DB - PRIME DP - Unbound Medicine ER -