Biochemical method for inserting new genetic information into DNA of Simian Virus 40: circular SV40 DNA molecules containing lambda phage genes and the galactose operon of Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 69 ·No. 10 ·1972-10-00 ·Pages 2904-9

Jackson DA, Symons RH, Berg P

Abstract

We have developed methods for covalently joining duplex DNA molecules to one another and have used these techniques to construct circular dimers of SV40 DNA and to insert a DNA segment containing lambda phage genes and the galactose operon of E. coli into SV40 DNA. The method involves: (a) converting circular SV40 DNA to a linear form, (b) adding single-stranded homodeoxypolymeric extensions of defined composition and length to the 3' ends of one of the DNA strands with the enzyme terminal deoxynucleotidyl transferase (c) adding complementary homodeoxypolymeric extensions to the other DNA strand, (d) annealing the two DNA molecules to form a circular duplex structure, and (e) filling the gaps and sealing nicks in this structure with E. coli DNA polymerase and DNA ligase to form a covalently closed-circular DNA molecule.

MeSH Terms

Centrifugation, Density Gradient Coliphages/enzymology,metabolism DNA Nucleotidyltransferases/metabolism DNA, Single-Stranded/metabolism DNA, Viral/analysis,metabolism Endonucleases/metabolism Escherichia coli/enzymology,metabolism Exonucleases/metabolism Galactose/biosynthesis Genetic Code Models, Structural Nucleic Acid Conformation Nucleic Acid Hybridization Operon Polynucleotide Ligases/metabolism Simian virus 40/metabolism Transformation, Genetic

Chemicals

DNA, Single-Stranded DNA, Viral DNA Nucleotidyltransferases Endonucleases Exonucleases Polynucleotide Ligases Galactose

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Jackson D A

Symons R H

Berg P

References (14)

14 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.69.10.2904

Published

1972-10-00

Pages

2904-9

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC389671

Subset

External Links

PubMed source DOI: 10.1073/pnas.69.10.2904 PMC Full Text View journal details

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Biochemical method for inserting new genetic information into DNA of Simian Virus 40: circular SV40 DNA molecules containing lambda phage genes and the galactose operon of Escherichia coli.

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