通过对需要丙氨酸的L5178Y细胞进行回复突变来分离营养缺陷型突变体和定量突变分析。

Isolation of nutrient deficient mutants and quantitative mutation assay by reversion of alanine-requiring L5178Y cells.

作者信息

Suzuki N, Okada S

出版信息

Mutat Res. 1976 Mar;34(3):489-505. doi: 10.1016/0027-5107(76)90225-6.

Abstract

Nutrient deficient cells for alanine or purine were isolated from L5178Y cells by the BudR-light method of Puck and Kao. The purine requiring cells, named P, showed maximum-plating efficiency in the presence of 10(-3)-10(-4) M inosine. The optimum concentration of L-alpha-alanine for Ala 32, one of alanine requiring mutants, was 10(-3) M. When Ala 32 cells were depleted of alanine, they showed an immediate decrease in incorporation of protein and DNA precursors without much change in that of RNA precursors and they ceased to multiply. Ala 32 cells have been used for experiments and have been phenotypically stable for 4 years. A quantitative mutation assay system for the reversion of L5178Y-Ala 32 cells from auxotrophy to prototrophy was established. The system was applied to some known mutagens, MNNG, 4-NQO, UV and gamma-rays. Some characteristics of the system are discussed and compared to drug-resistant mutation systems.

摘要

通过帕克（Puck）和考（Kao）的5-溴脱氧尿苷-光照法从L5178Y细胞中分离出丙氨酸或嘌呤营养缺陷型细胞。需要嘌呤的细胞，命名为P，在10⁻³ - 10⁻⁴ M肌苷存在下表现出最大铺板效率。对于丙氨酸需求突变体之一的Ala 32，L-α-丙氨酸的最佳浓度为10⁻³ M。当Ala 32细胞耗尽丙氨酸时，它们显示出蛋白质和DNA前体掺入立即减少，而RNA前体的掺入没有太大变化，并且它们停止增殖。Ala 32细胞已用于实验，并且在表型上稳定了4年。建立了一种用于将L5178Y-Ala 32细胞从营养缺陷型回复到原养型的定量突变检测系统。该系统应用于一些已知的诱变剂，如N-甲基-N'-硝基-N-亚硝基胍（MNNG）、4-硝基喹啉-1-氧化物（4-NQO）、紫外线（UV）和γ射线。讨论了该系统的一些特性，并与耐药突变系统进行了比较。