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- W2016862635 abstract "DNA polymerase from either Micrococcus luteus or Escherichia coli catalyzes the unprimed (de novo) incorporation of complementary deoxyribonucleoside triphosphates into double-stranded polynucleotides of defined repeating sequence. The de novo reactions of both enzymes, each tested at two levels of purification, show that subtle changes in reaction conditions (i.e. pH, buffer and metal ions) markedly influence the sequence of nucleotides in the polymeric product. Some of the results are: (a) with the partially purified M. luteus DNA polymerase in Tris-HCl buffer with the substrates dATP + dTTP, poly dA · poly dT is formed only under highly restricted conditions of pH (7.9 to 8.3); the alternating copolymer is found at all other pH ranges tested (7.3 to 7.9 and 8.3 to 8.7). Also, buffer ionic strength has a pronounced influence on these de novo reactions; at low concentrations of Tris-HCl buffer (15 mm, pH 8.1), the product is a mixture of poly dA · poly dT and poly d(A-T) · poly d(A-T). However, at high concentrations of Tris-HCl buffer (100 mm, pH 8.1), the product is all poly d(A-T) · poly d(A-T). The E. coli DNA polymerase synthesizes only poly d(A-T) · poly d(A-T) under a variety of conditions, (b) The E. coli DNA polymerase in the presence of dITP + dCTP synthesizes de novo either poly d(I-C) · poly d(I-C) or poly dI · poly dC depending on reaction conditions. The variables studied were buffer composition, buffer concentration, metal ion and reaction pH. Similar results are found with the highly purified M. luteus DNA polymerase in some reactions. (c) With dGTP + dCTP as substrates, only the partially purified E. coli DNA polymerase shows a de novo reaction; the product is poly dG · poly dC. None of the other enzyme preparations give a de novo reaction (within a 50-hr time period) with this set of substrates.Results indicate that the observed product specificity under different ionic conditions is not due to preferential degradation by the nucleases which are present in all enzyme preparations. Changes in the DNA polymerase molecule are apparently responsible for the observed product specificity.These studies are of interest since: (1) they may provide further insights into the mechanism of action of DNA polymerase; (2) they may relate to the natural occurrence of some of these DNA's; and (3) they are generally useful for the preparation of DNA's of defined and repeating nucleotide sequences." @default.
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- W2016862635 date "1964-05-01" @default.
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- W2016862635 title "The enzymatic synthesis of a copolymer of 6-methyl deoxyadenylate and deoxythymidylate" @default.
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- W2016862635 doi "https://doi.org/10.1016/0006-291x(64)90207-4" @default.
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