Construction of Biologically Functional Bacterial Plasmids In Vitro

The construction of new plasmid DNA species by in vitro joining of restriction endonuclease-generated fragments of separate plasmids is described. Newly constructed plasmids that are inserted into Escherichia coli by transformation are shown to be biologically functional replicons that possess genet...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1973-11, Vol.70 (11), p.3240-3244
Main Authors: Cohen, Stanley N., Annie C. Y. Chang, Boyer, Herbert W., Helling, Robert B.
Format: Article
Language:English
Subjects:
Base Sequence
Biological Sciences: Biochemistry
Centrifugation
Centrifugation, Density Gradient
Chloramphenicol
Circular DNA
DNA
DNA, Circular - isolation & purification
DNA, Viral - analysis
DNA, Viral - biosynthesis
DNA, Viral - isolation & purification
Drug Resistance, Microbial
Electrophoresis
Electrophoresis, Starch Gel
Endonucleases - metabolism
Escherichia coli - enzymology
Gels
Genetics, Microbial
Inclusion Bodies, Viral
Kanamycin
Molecular weight
Molecules
Nucleotides
Plasmids
Replicon
Tetracycline
Transformation, Genetic
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Summary:The construction of new plasmid DNA species by in vitro joining of restriction endonuclease-generated fragments of separate plasmids is described. Newly constructed plasmids that are inserted into Escherichia coli by transformation are shown to be biologically functional replicons that possess genetic properties and nucleotide base sequences from both of the parent DNA molecules. Functional plasmids can be obtained by reassociation of endonuclease-generated fragments of larger replicons, as well as by joining of plasmid DNA molecules of entirely different origins.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.70.11.3240