Suzuki M, Christians F C, Kim B, Skandalis A, Black M E, Loeb L A
Department of Pathology, Joseph Gottstein Memorial Cancer Research Laboratory, University of Washington, Seattle 98195-7705, USA.
Mol Divers. 1996 Oct;2(1-2):111-8. doi: 10.1007/BF01718708.
Random mutagenesis of genes followed by positive genetic selection in bacteria requires that the variant molecules confer biological activity, and is thus the most demanding approach for generating new functionally active molecules. Furthermore, one can learn much about the protein in question by comparing the population of selected molecules to the library from which they were selected. Described here is a mathematical method designed to guide such comparisons. We use as examples the results of randomization-selection studies of four different proteins. There exists, in general, a positive correlation between the number of amino acid substitutions in a critical region of a protein and the likelihood of inactivation of that protein; a correlation long suspected, but developed here in detail. At this time, we are comparing regions in different proteins and our conclusions must be limited. However, the method presented can serve as a guideline for anticipating the yield of new active mutants in genetic complementation assays based on the extent of randomization.
在细菌中对基因进行随机诱变，随后进行正向遗传筛选，这要求变异分子具有生物活性，因此这是生成新的功能活性分子最苛刻的方法。此外，通过将所选分子群体与其所来源的文库进行比较，可以了解很多关于所研究蛋白质的信息。这里描述的是一种旨在指导此类比较的数学方法。我们以四种不同蛋白质的随机化 - 筛选研究结果为例。一般来说，蛋白质关键区域的氨基酸取代数量与该蛋白质失活的可能性之间存在正相关；这种相关性早就被怀疑，但在此处进行了详细阐述。此时，我们正在比较不同蛋白质中的区域，我们的结论必然是有限的。然而，所提出的方法可以作为基于随机化程度预测基因互补试验中新活性突变体产量的指导原则。
