Siderovski D P, Mak T W
Amgen Institute, University of Toronto, Ontario, Canada.
Comput Biol Med. 1993 Nov;23(6):463-74. doi: 10.1016/0010-4825(93)90094-h.
Random mutagenesis is a powerful tool in protein structure-function analyses. One approach to random mutagenesis is the de novo synthesis of polypeptide-encoding oligodeoxy-nucleotides using doped nucleoside phosphoramidites. A Turbo PASCAL program, RAMHA, is described for modeling such mutagenesis. Upon entering the target sequence and the desired level of nucleotide contamination, RAMHA performs a Monte Carlo simulation of the mutagenesis, compiling statistics on the similarity of resultant mutant polypeptides to the wild-type sequence, the frequency of premature open-reading frame terminations, and other relevant outcomes. Simulated mutagenesis of two DNA targets has led to the development of two different strategies to avoid the random introduction of stop codons within mutagenized gene segments.
随机诱变是蛋白质结构-功能分析中的一种强大工具。随机诱变的一种方法是使用掺杂的核苷亚磷酰胺从头合成编码多肽的寡脱氧核苷酸。本文描述了一个用于模拟这种诱变的Turbo Pascal程序RAMHA。输入目标序列和所需的核苷酸污染水平后,RAMHA对诱变进行蒙特卡罗模拟,汇编有关所得突变多肽与野生型序列的相似性、过早开放阅读框终止的频率以及其他相关结果的统计数据。对两个DNA靶标的模拟诱变导致了两种不同策略的开发,以避免在诱变基因片段中随机引入终止密码子。