Deoxyribonucleic Acid Synthesis in Cell-free Extracts
Journal of Biological ChemistryPublished 1 September 1972Open access
Thomas B. Kornberg, Malcolm L. Gefter
Citations146
SJR quartileQ1
SJR score1.71
SNIP1.00
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TL;DR
DNA polymerase III, an enzyme essential for DNA replication, has been purified more than 10,000-fold from cell-free extracts of Escherichia coli and is present in amounts sufficient to account for the in vivo rate of replication.
Abstract
DNA polymerase III, an enzyme essential for DNA replication, has been purified more than 10,000-fold from cell-free extracts of Escherichia coli. The enzyme, judged to be 10 to 20% pure, requires all four deoxynucleoside 5′-triphosphates, Mg++, ethanol, and native DNA for maximal activity. It is present in amounts sufficient to account for the in vivo rate of replication. DNA polymerase III activity is inhibited by sulfhydryl inhibitors and by salt; it is not inhibited by antiserum directed against DNA polymerase I. The most active template for DNA polymerase III is DNA degraded partially by exonuclease III. Linear duplex DNA, single-stranded DNA, or DNA with single strand scissions is not used as template. The enzyme requires a primer to initiate synthesis and polymerization proceeds in the 5′ to 3′ direction by covalent extension of the primer.
Keywords
MedicineBiochemistry, Genetics and Molecular Biology
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