Replication of the Escherichia coli chromosome with a soluble enzyme system.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 71 ·No. 8 ·1974-08-00 ·Pages 3189-93

Kornberg T, Lockwood A, Worcel A

Abstract

Semi-conservative DNA synthesis is observed when the isolated, folded E. coli chromosome is supplemented with a DNA-free, soluble enzyme fraction, the four deoxynucleoside 5'-triphosphates, ATP, and Mg(++). The DNA synthesized in vitro remains associated with the folded chromosome during sedimentation through neutral sucrose, but is released as small DNA fragments in alkali. Sealing of these replicative intermediates to the chromosome requires the presence of both E. coli DNA polymerase I (EC 2.7.7.7) and DNA ligase (EC 6.5.1.2).

MeSH Terms

Adenosine Triphosphate/metabolism Carbon Radioisotopes Cell-Free System Centrifugation, Density Gradient Chromosomes, Bacterial/metabolism Cytosine Nucleotides/metabolism DNA Nucleotidyltransferases/metabolism DNA Replication DNA, Bacterial/biosynthesis Deoxyribonucleotides/metabolism Electrophoresis, Polyacrylamide Gel Escherichia coli/enzymology,metabolism Guanosine Triphosphate/metabolism Kinetics Nucleic Acid Conformation Polynucleotide Ligases/metabolism Protein Biosynthesis Templates, Genetic Thymine/metabolism Tritium Uracil Nucleotides/metabolism

Chemicals

Carbon Radioisotopes Cytosine Nucleotides DNA, Bacterial Deoxyribonucleotides Uracil Nucleotides Tritium Guanosine Triphosphate Adenosine Triphosphate DNA Nucleotidyltransferases Polynucleotide Ligases Thymine

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Kornberg T

Lockwood A

Worcel A

References (24)

24 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.71.8.3189

Published

1974-08-00

Pages

3189-93

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC388648

Subset

External Links

PubMed source DOI: 10.1073/pnas.71.8.3189 PMC Full Text View journal details

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Replication of the Escherichia coli chromosome with a soluble enzyme system.

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