Single-step selection of mouse FM3A cell mutants defective in thymidylate synthetase
Single-step selection of mouse FM3A cell mutants defective in thymidylate synthetase
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胸苷酸合成酶缺陷型小鼠 FM3A 细胞突变体的一步选择
DOI：
10.1007/bf01538800
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发表时间：
1980
期刊：
影响因子：
--
通讯作者：
T. Seno
中科院分区：
文献类型：
--
作者：
D. Ayusawa;H. Koyama;K. Iwata;T. Seno
介绍了一种用氚自杀法分离胸苷营养缺陷型突变体的方法。在含有[3 H]脱氧尿苷的培养基中培养诱变的小鼠FM 3A细胞。大多数存活的克隆检查显示绝对胸苷营养缺陷型的表型。这种表型非常稳定，并且在细胞-细胞杂交实验中被发现是遗传隐性的。这些变体克隆的生长不受除胸苷以外的各种嘧啶核苷的支持。这些克隆的粗提物中胸苷酸合成酶的活性不到FM 3A细胞的1%。这些结果有力地表明，胸苷营养缺陷型是由胸苷酸合成酶的遗传缺陷引起的。
A tritium-suicide method for isolating thymidine auxotrophic mutants is described. Mutagenized mouse FM3A cells were cultured in medium containing [3H] deoxyuridine. Most of the surviving clones examined showed a phenotype of absolute thymidine auxotrophy. This phenotype is very stable and was found to be genetically recessive in cell-cell hybridization experiments. The growth of these variant clones was not supported by various pyrimidine nucleosides other than thymidine. The activity of thymidylate synthetase in crude extracts of these clones was less than 1% of that of FM3A cells. These results strongly indicate that the thymidine auxotrophic phenotype resulted from a genetic defect in thymidylate synthetase.