Hinnebusch A G, Clark V E, Klotz L C
Biochemistry. 1978 Apr 18;17(8):1521-9. doi: 10.1021/bi00601a026.
The reassociation kinetics have been measured for radioactive Escherichia coli DNAs (tracers) of various average single-strand lengths reassociated alone and in the presence of excess unlabeled DNA (driver) of two different average lengths. Hydroxylapatite binding was used to follow the reaction time course. The length-dependence of the rate constant determined in the tracer self-reassociation reactions is in agreement with the square-root dependence previously determined (Wetmur, J. G., & Davisond, N. (1968) J. Mol. Biol. 31, 349-370) using optical methods to follow the time course. However, for the driver-tracer reactions, where the radioactive DNA reassociates largely with DNA of a different average length, the dependence of the rate constant upon average tracer length is increased and approaches an L to the first power dependence. In 0.18 M Na+, the variation of the rate constant for tracer reassociation with the lengths of the reassociating strands has been shown to fit the simple equation k = (9.0077).(L T 0.55 + 1/L D 0.55), where k is the observed rate constant in L mol-1 s-1 and L(T)and L(D) are the weight average tracer and driver lengths, respectively, in nucleotides. This dependence suggests that the rate of nucleation of two free strands is proportional to the sum of the reciprocals of the hydrodynamic radii of the two strands.
已测定了各种平均单链长度的放射性大肠杆菌DNA(示踪剂)在单独重退火以及在存在两种不同平均长度的过量未标记DNA(驱动子)的情况下的重退火动力学。使用羟基磷灰石结合来跟踪反应的时间进程。在示踪剂自重退火反应中测定的速率常数对长度的依赖性与先前使用光学方法跟踪时间进程所确定的平方根依赖性一致(Wetmur,J.G.,& Davisond,N.(1968)J. Mol. Biol. 31,349 - 370)。然而,对于驱动子 - 示踪剂反应,其中放射性DNA主要与不同平均长度的DNA重退火,速率常数对平均示踪剂长度的依赖性增加,并接近一次幂的L依赖性。在0.18 M Na⁺中,示踪剂重退火的速率常数随重退火链长度的变化已显示符合简单方程k = (9.0077)·(L(T)⁰.⁵⁵ + 1/L(D)⁰.⁵⁵),其中k是以L mol⁻¹ s⁻¹为单位的观测速率常数,L(T)和L(D)分别是以核苷酸为单位的重均示踪剂和驱动子长度。这种依赖性表明两条自由链的成核速率与两条链的流体动力学半径的倒数之和成正比。