Chernov A P, Ivanov V A
Biokhimiia. 1995 Jun;60(6):874-82.
Preparations of Rous sarcoma virus reverse transcriptase isolated from a culture of E. coli HB101 (pMF14) and purified to homogeneity were used to study the steady state kinetics of DNA polymerization and inhibition of DNA-polymerase activity. DNA synthesis was examined using a system of poly(rA) as template, oligo(dT) as primer and dTTP as nucleotide substrate. Kinetic constants for steady state conditions were determined. The substrate initial velocity patterns point to an ordered mechanism which results in the formation of a ternary complex, in which the template-primer is the first to bind to the enzyme. Inhibition of the DNA-polymerase activity of the enzyme by various inhibitors was studied. Analysis of final products of the DNA-polymerase reaction revealed the presence of distribution syntheses of the DNA chain by the alpha alpha-subunit form of the enzyme.
从大肠杆菌HB101（pMF14）培养物中分离并纯化至同质的劳氏肉瘤病毒逆转录酶制剂，用于研究DNA聚合的稳态动力学以及DNA聚合酶活性的抑制作用。使用聚（rA）作为模板、寡聚（dT）作为引物和dTTP作为核苷酸底物的系统来检测DNA合成。测定了稳态条件下的动力学常数。底物初始速度模式表明存在一种有序机制，该机制导致形成三元复合物，其中模板 - 引物首先与酶结合。研究了各种抑制剂对该酶DNA聚合酶活性的抑制作用。对DNA聚合酶反应最终产物的分析表明，该酶的αα亚基形式存在DNA链的分布合成。
