Tan C K, So M J, Downey K M, So A G
Nucleic Acids Res. 1987 Mar 11;15(5):2269-78. doi: 10.1093/nar/15.5.2269.
The mechanism by which millimolar concentrations of ATP stimulate the activity and increase the processivity of calf thymus DNA polymerase alpha has been investigated with poly(dA)/oligo(dT) as template/primer to eliminate possible effects due to primer synthesis. The effect of ATP on the rate of DNA synthesis with this template/primer was found to be dependent upon whether or not the ATP was neutralized and the species of buffer used in the reaction. The present studies suggest that ATP stimulation of calf thymus DNA polymerase can be attributed to changes in the pH of the reaction mixture, a shift in the magnesium ion optimum, or both. Furthermore, effects of ATP on the processivity of DNA polymerase alpha could be mimicked by lowering the pH of the reaction mixture.
以聚(dA)/寡聚(dT)作为模板/引物,研究了毫摩尔浓度的ATP刺激小牛胸腺DNA聚合酶α活性并提高其持续合成能力的机制,以消除引物合成可能产生的影响。发现ATP对该模板/引物的DNA合成速率的影响取决于ATP是否被中和以及反应中使用的缓冲液种类。目前的研究表明,ATP对小牛胸腺DNA聚合酶的刺激作用可归因于反应混合物pH值的变化、镁离子最佳浓度的改变或两者兼而有之。此外,降低反应混合物的pH值可以模拟ATP对DNA聚合酶α持续合成能力的影响。