Bauer G J, McCaskill J S, Otten H
Max-Planck-Institut für Biophysikalische Chemie, Göttingen, Federal Republic of Germany.
Proc Natl Acad Sci U S A. 1989 Oct;86(20):7937-41. doi: 10.1073/pnas.86.20.7937.
Populations of short self-replicating RNA variants have been confined to one side of a reaction-diffusion traveling wave front propagating along thin capillary tubes containing the Q beta viral enzyme. The propagation speed is accurately measurable with a magnitude of about 1 micron/sec, and the wave persists for hundreds of generations (of duration less than 1 min). Evolution of RNA occurs in the wavefront, as established by front velocity changes and gel electrophoresis of samples drawn from along the capillary. The high population numbers (approximately equal to 10(11], their well-characterized biochemistry, their short generation time, and the constant conditions make the system ideal for evolution experiments. Growth is monitored continuously by excitation of an added RNA-sensitive fluorescent dye, ethidium bromide. An analytic expression for the front velocity is derived for the multicomponent kinetic scheme that reduces, for a high RNA-enzyme binding constant, to the Fisher form v = 2 square root of kappa D, where D is the diffusion constant of the complex and kappa is the low-concentration overall replication rate coefficient. The latter is confirmed as the selective value-determining parameter by numerical solution of a two-species system.
短的自我复制RNA变体群体被限制在沿着含有Qβ病毒酶的细毛细管传播的反应扩散行波前沿的一侧。传播速度可精确测量,大小约为1微米/秒,并且波持续数百代(持续时间小于1分钟)。如通过前沿速度变化以及从毛细管沿线抽取的样品的凝胶电泳所确定的,RNA的进化发生在波前。高群体数量(约等于10¹¹)、其特征明确的生物化学性质、其短的世代时间以及恒定的条件使该系统成为进化实验的理想选择。通过添加的对RNA敏感的荧光染料溴化乙锭的激发来连续监测生长。针对多组分动力学方案推导了前沿速度的解析表达式,对于高RNA - 酶结合常数,该表达式简化为费舍尔形式v = 2√κD,其中D是复合物的扩散常数,κ是低浓度下的总体复制速率系数。通过双物种系统的数值解证实后者是选择性值决定参数。