Abstract
Construction and characterization of a class of multicopy plasmid cloning vehicles containing the replication system of miniplasmid P15A are described. The constructed plasmids have cleavage sites within antibiotic resistance genes for a variety of commonly employed site-specific endonucleases, permitting convenient use of the insertional inactivation procedure for the selection of clones that contain hybrid DNA molecules. Although the constructed plasmids showed DNA sequence homology with the ColE1 plasmid within the replication region, were amplifiable by chloramphenicol or spectinomycin, required DNA polymerase I for replication, and shared other replication properties with ColE1, they were nevertheless compatible with ColE1. P15A-derived plasmids were not self-transmissible and were mobilized poorly by Hfr strains; however, mobilization was complemented by the presence of a ColE1 plasmid within the same cell.
Keywords
ColE1BiologyPlasmidT-DNA Binary systemRepliconMultiple cloning siteGeneticsMolecular cloningMolecular biologyPlasmid preparationDNA replicationCloning vectorDNAOrigin of replicationGeneRecombinant DNAPBR322Vector (molecular biology)Peptide sequence
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Publication Info
- Year
- 1978
- Type
- article
- Volume
- 134
- Issue
- 3
- Pages
- 1141-1156
- Citations
- 4268
- Access
- Closed
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Cite This
Annie Chang,
Stanley N. Cohen
(1978).
Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid.
Journal of Bacteriology
, 134
(3)
, 1141-1156.
https://doi.org/10.1128/jb.134.3.1141-1156.1978
Identifiers
- DOI
- 10.1128/jb.134.3.1141-1156.1978