禽成髓细胞瘤病毒DNA聚合酶进行DNA合成的模板特异性条件观察
Observations on template-specific conditions for DNA synthesis by avian myeloblastosis virus DNA polymerase.
作者信息
Marcus S L, Modak M J
出版信息
Nucleic Acids Res. 1976 Jun;3(6):1473-86. doi: 10.1093/nar/3.6.1473.
Abstract
The effects of Mg++, Mn++, and KCl addition, individually and in combination, on the rate of DNA- and RNA-primed DNA synthesis by avian myeloblastosis virus DNA polymerase (reverse transcriptase) using a variety of natural and synthetic template-primer combinations were examined. Optimal divalent cation concentrations were found to vary by as much as 10-fold depending upon the template-primer used to direct synthesis. Addition of KCl to reaction mixtures containing optimal divalent cation concentrations produced stimulation or inhibition of DNA synthesis which was also template-specific. DNA synthesis on the modified template poly (2'-0-methylcytidylate) was uniquely stimulated by combinations of divalent cations. With Mg++ as divalent cation, deviations from classical Michaelis-Menten kinetics of substrate saturation were observed with all template-primers tested.
摘要
研究了单独添加和组合添加Mg++、Mn++和KCl对禽成髓细胞瘤病毒DNA聚合酶（逆转录酶）利用多种天然和合成模板-引物组合进行DNA引发的DNA合成以及RNA引发的DNA合成速率的影响。发现最佳二价阳离子浓度根据用于指导合成的模板-引物不同而变化高达10倍。向含有最佳二价阳离子浓度的反应混合物中添加KCl会产生对DNA合成的刺激或抑制，这也是模板特异性的。在修饰模板聚（2'-O-甲基胞苷酸）上的DNA合成受到二价阳离子组合的独特刺激。以Mg++作为二价阳离子时，在所测试的所有模板-引物中均观察到与经典米氏底物饱和动力学的偏差。
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