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PMID: 1094460 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Cloning, isolation, and characterization of replication regions of complex plasmid genomes.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 72 ·No. 6 ·1975-06-00 ·Pages 2242-6

Timmis K, Cabello F, Cohen SN

Abstract

EcoRI endonuclease-generated DNA fragments carrying replication regions of the F'lac and R6-5 plasmids have been cloned and isolated, using as a selection vehicle a nonreplicating ampicillin-resistance DNA fragment derived from a Staphylococcus aureus plasmid. Heteroduplex analysis of the constructed plasmid chimeras and the parent replicons has localized the cloned R6-5 replication region to a DNA segment between kilobase pair coordinates 1.0 and 88.0 on the R6-5 map. Physical proximity between the plasmid replication functions and the locus governing plasmid incompatibility has been shown for both parent replicons. The cloning method reported appears to be generally applicable for the identification and isolation of replication regions of a variety of complex genomes.

MeSH Terms
Centrifugation, Density Gradient DNA Replication DNA Restriction Enzymes DNA, Bacterial/isolation & purification Electrophoresis, Agar Gel Escherichia coli/analysis,ultrastructure Extrachromosomal Inheritance Genetics, Microbial Hybridization, Genetic Microscopy, Electron Molecular Weight Phenotype Plasmids Selection, Genetic Staphylococcus Transformation, Genetic
Chemicals
DNA, Bacterial DNA Restriction Enzymes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Timmis K
Cabello F
Cohen S N
References (28)
28 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.72.6.2242
Published
1975-06-00
Pages
2242-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC432733
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.72.6.2242 PMC Full Text View journal details
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