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The mitochondrial DNA (mtDNA) polymerase was isolated from a protease-deficient yeast strain (PY2), and purified about 3000 fold by a column chromatography on phosphocellulose, heparin-agarose, and single-stranded DNA cellulose. The purified polymerase was characterized with respect to optimal nucleotide concentration, template-primer specificity and sensitivity to some inhibitors. These results were compared with the nuclear DNA polymerase I activity. Both polymerases showed similar requirement of deoxynucleotide concentrations (Km < 1 uM), and highest activity with poly(dA-dT) template. However, the mtDNA polymerase was more sensitive to ddTTP, EtBr and Mn2+ inhibition in comparison to the nuclear DNA polymerase I. The mtDNA polymerase did not need ATP as an energy source for in vitro DNA synthesis. This mtDNA polymerase preparation also showed 3' -> 5' exonudease activity.
The purpose of present study is to investigate the efficiency of different kinds of Turkish commercial bleaching earth materials for changes in different colour pigment concentrations in neutralized sunfl ower oils. The bleaching experiments were performed in a pilot system under at stable vacuum (50 mmHg) and temperature (100ºC) for 30 min. By examining the changes in chlorophyll, β-carotene and red colour, bleaching process parameters such as type and dosage of the bleaching material were optimised. The sorption characteristics of colour pigments were evaluated using common adsorption isotherms and Scatchard plot analysis. Ads-3 acid-activated earth material at 1% (w/w) per samples was found to be the most appropriate sorbent and the amount of sorbed pigments was calculated as 1.01x10–4 mmol/g ads. for chlorophyll, 1.15x10–3 mmol/g ads. for carotene and 1.70 red on Lovibond colour scale. The procedure indicated that this system can be easily adapted to the actual oil refi ning systems.
Mitochondrial DNA polymerase from Saccharomyces cerevisiae, purified 3500 fold, was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis into three polypeptides. The major 150 kDa polypeptide was probably the catalytic subunit of the mitochondrial (mt) DNA polymerase and the other two polypeptides could be either proteolytic cleavage products of the polymerase, other subunits of the enzyme or protein contaminants. The mtDNA polymerase preferred an A+T-rich DNA template and did not require any RNA primer for DNA synthesis, at least under in vitro reaction conditions. It showed higher processivity on a double-stranded linear DNA template than on a single-stranded circular DNA template, and was capable of synthesizing at least about 1200 nucleotide primer-extended products without any major pause on a double-stranded DNA template.
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