Partial purification of a chloroplast DNA polymerase from Euglena gracilis |
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Authors: | S J Keller S A Biedenbach R R Meyer |
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Affiliation: | Department of Biological Sciences, University of Cincinnati Cincinnati, Ohio 45221 USA |
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Abstract: | A single DNA polymerase has been purified 965 fold from isolated chloroplasts of with a yield of 53%. The isolation methods include solubilization of the enzyme with 1M NaCl, ammonium sulfate precipitation, DNA affinity and DEAE-cellulose chromatography. The enzyme requires all four deoxynucleotide triphosphates, magnesium and denatured DNA for maximal activity. The chloroplast DNA polymerase is free of contaminating nucleases and phosphatases, has a sharp pH optimum at pH 7.2 and magnesium optimum of 6mM. |
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