Isolation and characterization of glucose-6-phosphate dehydrogenase-deficient Chinese hamster cells derived from pure mutant colonies.
Isolation and characterization of glucose-6-phosphate dehydrogenase-deficient Chinese hamster cells derived from pure mutant colonies.
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来自纯突变体菌落的葡萄糖-6-磷酸脱氢酶缺陷型中国仓鼠细胞的分离和表征。
DOI：
10.1093/mutage/4.4.259
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发表时间：
1989
期刊：
影响因子：
2.7
通讯作者：
Giaccia,A
中科院分区：
文献类型：
--
作者：
Stamato,TD;Richardson,E;Ianacone,J;MacLaren,RA;Denko,N;Giaccia,A
从完全由缺乏G6 PD染色活性的细胞组成的集落中分离出10个纯的葡萄糖-6-磷酸脱氢酶（G6 PD）缺陷型突变体。这些突变体进行了分析G6 PD酶活性和免疫交叉反应蛋白的存在下，使用免疫印迹技术和抗血清针对牛G6 PD。四个突变体没有可检测的酶活性，并且不含有与G6 PD抗体产生可检测的交叉反应的蛋白质。一个突变体有残留的酶活性和改变电泳迁移率，但没有检测到免疫交叉反应。这些结果可以通过DNA缺失或点突变机制来解释。另一方面，分析的10个突变体中有5个具有与G6 PD基因中的点突变一致的特征。所有这些都含有一种与G6 PD抗体交叉反应的蛋白质，并且具有与亲本细胞的G6 PD酶相同的亚基分子量。五个突变体中的四个具有残留的G6 PD酶活性。因此，形成纯突变体的机制并不简单是DNA缺失，而可能是一个更复杂的过程，涉及改变的遗传信息从一条DNA链转移到另一条DNA链。
Ten pure glucose-6-phosphate dehydrogenase (G6PD)-deficient mutants were isolated from colonies composed entirely of cells which lacked G6PD staining activity. These mutants were analyzed for G6PD enzyme activity and the presence of immunologically cross-reactive proteins using immunoblotting techniques and antiserum directed against bovine G6PD. Four mutants had no detectable enzyme activity and did not contain protein which produces a detectable cross-reaction with G6PD antibody. One mutant had residual enzyme activity and altered electrophoretic mobility but did not have detectable immunological crossreactivity. These results could be explained by either a DNA deletion or point mutational mechanism. On the other hand, five of the 10 mutants analyzed had characteristics consistent with a point mutation in the G6PD gene. All contain a protein which cross-reacts with the G6PD antibody and have the same subunit molecular weight as the parent cell's G6PD enzyme. Four of the five mutants had residual G6PD enzyme activity. Thus, the mechanism for the formation of pure mutants is not simply DNA deletion but is probably a more complex process involving the transfer of altered genetic information from one DNA strand to the other.