Mutagenesis during in vitro DNA synthesis.

Abstract:

:The error frequency of in vitro DNA synthesis using a natural DNA template has been measured with a biological assay for nucleotide substitutions. phiX174 DNA containing an amber mutation was copied in vitro by Escherichia coli DNA polymerase I, and the reversion frequency of the progeny DNA was determined by transfection of E. coli spheroplasts. E. coli polymerase I makes less than 1 mistake at the am3 locus for every 7700 nucleotides incorporated under standard reaction conditions. Substitution of Mn2+ for Mg2+ and unequal concentrations of deoxynucleoside triphosphate substrates raises this mutation frequency to greater than 1 in 1000. Thus, E. coli DNA polymerase I can copy natural DNA templates with high fidelity and its accuracy can be affected by alterations in reaction conditions.

authors

Weymouth LA,Loeb LA

doi

10.1073/pnas.75.4.1924

subject

Has Abstract

pub_date

1978-04-01 00:00:00

pages

1924-8

issue

4

eissn

0027-8424

issn

1091-6490

journal_volume

75

pub_type

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