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Replication defective RP4 plasmids recovered after chromosomal integration.
Plasmid. 1984 Jan; 11(1):65-73.P

Abstract

pHH6000 is a composite replicon made by the in vitro ligation of the IncP plasmid RP4 to a fragment of bacteriophage lambda capable of autonomous replication. Derivatives were selected in which it had integrated into the Escherichia coli chromosome by homologous recombination with the resident lambda prophage, and plasmids were subsequently regenerated from the integrated molecules. Although of the same molecular size as pHH6000, all had altered properties: those recovered from the chromosome of cells simultaneously carrying a distinguishable autonomous IncP plasmid showed a 100- to 1000-fold reduction in their ability to become established in a lambda lysogen; those regenerated from cells with no autonomous IncP plasmid were no longer RP4 replicons, now being dependent on replication functions encoded by the lambda DNA they carry and therefore unable to form a plasmid in a lambda lysogen. This second class of plasmids still exhibited normal RP4 incompatibility and stability even though neither property is encoded by the lambda replicator DNA. It was concluded that expression of RP4 incompatibility and partitioning control do not require an intact RP4 replicon. The data also suggest that the presence in the chromosome of a normal RP4 molecule may be deleterious to the host, although the manner in which the integrated molecules were obtained allows other explanations. The composite plasmids replicating from cloned lambda genes should be useful in analysis of the regulated distribution of RP4 molecules at cell division.

Authors

No affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

6231650

Citation

Grinter, N J.. "Replication Defective RP4 Plasmids Recovered After Chromosomal Integration." Plasmid, vol. 11, no. 1, 1984, pp. 65-73.
Grinter NJ. Replication defective RP4 plasmids recovered after chromosomal integration. Plasmid. 1984;11(1):65-73.
Grinter, N. J. (1984). Replication defective RP4 plasmids recovered after chromosomal integration. Plasmid, 11(1), 65-73.
Grinter NJ. Replication Defective RP4 Plasmids Recovered After Chromosomal Integration. Plasmid. 1984;11(1):65-73. PubMed PMID: 6231650.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Replication defective RP4 plasmids recovered after chromosomal integration. A1 - Grinter,N J, PY - 1984/1/1/pubmed PY - 1984/1/1/medline PY - 1984/1/1/entrez SP - 65 EP - 73 JF - Plasmid JO - Plasmid VL - 11 IS - 1 N2 - pHH6000 is a composite replicon made by the in vitro ligation of the IncP plasmid RP4 to a fragment of bacteriophage lambda capable of autonomous replication. Derivatives were selected in which it had integrated into the Escherichia coli chromosome by homologous recombination with the resident lambda prophage, and plasmids were subsequently regenerated from the integrated molecules. Although of the same molecular size as pHH6000, all had altered properties: those recovered from the chromosome of cells simultaneously carrying a distinguishable autonomous IncP plasmid showed a 100- to 1000-fold reduction in their ability to become established in a lambda lysogen; those regenerated from cells with no autonomous IncP plasmid were no longer RP4 replicons, now being dependent on replication functions encoded by the lambda DNA they carry and therefore unable to form a plasmid in a lambda lysogen. This second class of plasmids still exhibited normal RP4 incompatibility and stability even though neither property is encoded by the lambda replicator DNA. It was concluded that expression of RP4 incompatibility and partitioning control do not require an intact RP4 replicon. The data also suggest that the presence in the chromosome of a normal RP4 molecule may be deleterious to the host, although the manner in which the integrated molecules were obtained allows other explanations. The composite plasmids replicating from cloned lambda genes should be useful in analysis of the regulated distribution of RP4 molecules at cell division. SN - 0147-619X UR - https://www.unboundmedicine.com/medline/citation/6231650/Replication_defective_RP4_plasmids_recovered_after_chromosomal_integration_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0147-619X(84)90008-8 DB - PRIME DP - Unbound Medicine ER -