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Isolation of the replication DNA region from a Rhizobium plasmid and examination of its potential as a replicon for Rhizobiaceae cloning vectors.
Plasmid 1990; 23(3):201-15P

Abstract

The DNA region essential for replication and stability of a native plasmid (pTM5) from Rhizobium sp. (Hedysarum) has been identified and isolated within a 5.4-kb PstI restriction fragment. The isolation of this region was accomplished by cloning endonuclease-restricted pTM5 DNA into a ColE1-type replicon and selecting the recombinant plasmids containing the pTM5 replicator (pTM5 derivative plasmids) by their ability to replicate in Rhizobium. DNA homology studies revealed that pTM5-like replicons are present in cryptic plasmids from some Rhizobium sp. (Hedysarum) strains but not in plasmids from strains of other Rhizobium species or Agrobacterium tumefaciens. The pTM5 derivative plasmids were able to replicate in Escherichia coli and A. tumefaciens and in a wide range of Rhizobium species. On the basis of stability assays in the absence of antibiotic selective pressure, the pTM5 derivative plasmids were shown to be highly stable in both free-living and symbiotic cells of Rhizobium sp. (Hedysarum). The stability of these plasmids in other species of Rhizobium and in A. tumefaciens varied depending on the host and on the plasmid. Most pTM5 derivative plasmids tested showed significantly higher symbiotic stability than RK2 derivative plasmids pRK290 and pAL618 in Rhizobium sp. (Hedysarum), R. meliloti, and R. leguminosarum by. phaseoli. Consequently, we consider that the constructed pTM5 derivative plasmids are potentially useful as cloning vectors for Rhizobiaceae.

Authors+Show Affiliations

Laboratorio de Microbiología, ETS Ingenieros Agrónomos, Universidad Politécnica, Madrid, Spain.No affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

2217572

Citation

Mozo, T, et al. "Isolation of the Replication DNA Region From a Rhizobium Plasmid and Examination of Its Potential as a Replicon for Rhizobiaceae Cloning Vectors." Plasmid, vol. 23, no. 3, 1990, pp. 201-15.
Mozo T, Cabrera E, Ruiz-Argüeso T. Isolation of the replication DNA region from a Rhizobium plasmid and examination of its potential as a replicon for Rhizobiaceae cloning vectors. Plasmid. 1990;23(3):201-15.
Mozo, T., Cabrera, E., & Ruiz-Argüeso, T. (1990). Isolation of the replication DNA region from a Rhizobium plasmid and examination of its potential as a replicon for Rhizobiaceae cloning vectors. Plasmid, 23(3), pp. 201-15.
Mozo T, Cabrera E, Ruiz-Argüeso T. Isolation of the Replication DNA Region From a Rhizobium Plasmid and Examination of Its Potential as a Replicon for Rhizobiaceae Cloning Vectors. Plasmid. 1990;23(3):201-15. PubMed PMID: 2217572.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Isolation of the replication DNA region from a Rhizobium plasmid and examination of its potential as a replicon for Rhizobiaceae cloning vectors. AU - Mozo,T, AU - Cabrera,E, AU - Ruiz-Argüeso,T, PY - 1990/5/1/pubmed PY - 1990/5/1/medline PY - 1990/5/1/entrez SP - 201 EP - 15 JF - Plasmid JO - Plasmid VL - 23 IS - 3 N2 - The DNA region essential for replication and stability of a native plasmid (pTM5) from Rhizobium sp. (Hedysarum) has been identified and isolated within a 5.4-kb PstI restriction fragment. The isolation of this region was accomplished by cloning endonuclease-restricted pTM5 DNA into a ColE1-type replicon and selecting the recombinant plasmids containing the pTM5 replicator (pTM5 derivative plasmids) by their ability to replicate in Rhizobium. DNA homology studies revealed that pTM5-like replicons are present in cryptic plasmids from some Rhizobium sp. (Hedysarum) strains but not in plasmids from strains of other Rhizobium species or Agrobacterium tumefaciens. The pTM5 derivative plasmids were able to replicate in Escherichia coli and A. tumefaciens and in a wide range of Rhizobium species. On the basis of stability assays in the absence of antibiotic selective pressure, the pTM5 derivative plasmids were shown to be highly stable in both free-living and symbiotic cells of Rhizobium sp. (Hedysarum). The stability of these plasmids in other species of Rhizobium and in A. tumefaciens varied depending on the host and on the plasmid. Most pTM5 derivative plasmids tested showed significantly higher symbiotic stability than RK2 derivative plasmids pRK290 and pAL618 in Rhizobium sp. (Hedysarum), R. meliloti, and R. leguminosarum by. phaseoli. Consequently, we consider that the constructed pTM5 derivative plasmids are potentially useful as cloning vectors for Rhizobiaceae. SN - 0147-619X UR - https://www.unboundmedicine.com/medline/citation/2217572/Isolation_of_the_replication_DNA_region_from_a_Rhizobium_plasmid_and_examination_of_its_potential_as_a_replicon_for_Rhizobiaceae_cloning_vectors_ DB - PRIME DP - Unbound Medicine ER -