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Construction of a novel plasmid-phage hybrid: use of the hybrid to demonstrate ColE1 DNA replication in vivo in the absence of a ColE1-specified protein.
Proc Natl Acad Sci U S A. 1978 May; 75(5):2200-4.PN

Abstract

A hybrid bacteriophage, P420, was constructed in vitro; it contains part of bacteriophage P4 and a 3.6-kilobase derivative of plasmid ColE1. In Escherichia coli, the plasmid-phage hybrid can exist as a stable plasmid or can be packaged into infective bacteriophage particles. Replication of P420, directed by the ColE1 replicon, was found to occur after P420 phage infection of E. coli cells that had been incubated in the presence of chloramphenicol. Replication began without a lag period and resulted in the synthesis of covalently closed circles of P420 DNA. Like ColE1 DNA replication but unlike that of P4, replication was dependent on DNA polymerase I and was sensitive to rifampicin. The presence of a resident ColE1 plasmid in the infected cells resulted in an inhibition of the replication of the incoming P420 DNA. These results indicate that ColE1 does not require a plasmid-coded protein to replicate its DNA in vivo and demonstrate the utility of P4 bacteriophage for coupling bacteriophage properties to a plasmid replicon.

Authors

No affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

276860

Citation

Kahn, M, and D R. Helinski. "Construction of a Novel Plasmid-phage Hybrid: Use of the Hybrid to Demonstrate ColE1 DNA Replication in Vivo in the Absence of a ColE1-specified Protein." Proceedings of the National Academy of Sciences of the United States of America, vol. 75, no. 5, 1978, pp. 2200-4.
Kahn M, Helinski DR. Construction of a novel plasmid-phage hybrid: use of the hybrid to demonstrate ColE1 DNA replication in vivo in the absence of a ColE1-specified protein. Proc Natl Acad Sci USA. 1978;75(5):2200-4.
Kahn, M., & Helinski, D. R. (1978). Construction of a novel plasmid-phage hybrid: use of the hybrid to demonstrate ColE1 DNA replication in vivo in the absence of a ColE1-specified protein. Proceedings of the National Academy of Sciences of the United States of America, 75(5), 2200-4.
Kahn M, Helinski DR. Construction of a Novel Plasmid-phage Hybrid: Use of the Hybrid to Demonstrate ColE1 DNA Replication in Vivo in the Absence of a ColE1-specified Protein. Proc Natl Acad Sci USA. 1978;75(5):2200-4. PubMed PMID: 276860.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Construction of a novel plasmid-phage hybrid: use of the hybrid to demonstrate ColE1 DNA replication in vivo in the absence of a ColE1-specified protein. AU - Kahn,M, AU - Helinski,D R, PY - 1978/5/1/pubmed PY - 1978/5/1/medline PY - 1978/5/1/entrez SP - 2200 EP - 4 JF - Proceedings of the National Academy of Sciences of the United States of America JO - Proc. Natl. Acad. Sci. U.S.A. VL - 75 IS - 5 N2 - A hybrid bacteriophage, P420, was constructed in vitro; it contains part of bacteriophage P4 and a 3.6-kilobase derivative of plasmid ColE1. In Escherichia coli, the plasmid-phage hybrid can exist as a stable plasmid or can be packaged into infective bacteriophage particles. Replication of P420, directed by the ColE1 replicon, was found to occur after P420 phage infection of E. coli cells that had been incubated in the presence of chloramphenicol. Replication began without a lag period and resulted in the synthesis of covalently closed circles of P420 DNA. Like ColE1 DNA replication but unlike that of P4, replication was dependent on DNA polymerase I and was sensitive to rifampicin. The presence of a resident ColE1 plasmid in the infected cells resulted in an inhibition of the replication of the incoming P420 DNA. These results indicate that ColE1 does not require a plasmid-coded protein to replicate its DNA in vivo and demonstrate the utility of P4 bacteriophage for coupling bacteriophage properties to a plasmid replicon. SN - 0027-8424 UR - https://www.unboundmedicine.com/medline/citation/276860/Construction_of_a_novel_plasmid_phage_hybrid:_use_of_the_hybrid_to_demonstrate_ColE1_DNA_replication_in_vivo_in_the_absence_of_a_ColE1_specified_protein_ L2 - https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/276860/ DB - PRIME DP - Unbound Medicine ER -