一种抗抗霉素A的人类细胞系的分离

The isolation of an antimycin A-resistant human cell line.

作者信息

Munro E, Webb M, Kearsey S E, Craig I W

出版信息

Exp Cell Res. 1983 Sep;147(2):329-39. doi: 10.1016/0014-4827(83)90215-x.

Abstract

An antimycin A-resistant derivative of the human cell line, D98, has been obtained by selective mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine. The derivative, designed MA65, is capable of continuous growth in 15 microM antimycin and the resistant phenotype is stable in the absence of selection. MA65 is not cross-resistant to chloramphenicol or triethyl tin. Crude membrane preparations from MA65 after propagation in medium containing antimycin have normal succinate-cytochrome c oxidoreductase activity and the respiratory activity of whole cells continues in the presence of the drug. The mitochondrially synthesized proteins of D98 and MA65 are similar when compared on sodium dodecyl sulphate (SDS), or isoelectric focusing gels, but there is a reproducible difference in the extent of labelling of one band detected by isoelectric focusing. Genetic analysis is consistent with the existence of a cytoplasmically localized determinant conferring resistance.

摘要

通过用N-甲基-N'-硝基-N-亚硝基胍进行选择性诱变，已获得人细胞系D98的抗抗霉素A衍生物。该衍生物命名为MA65，能够在15微摩尔抗霉素中持续生长，并且在没有选择压力的情况下抗性表型稳定。MA65对氯霉素或三乙基锡无交叉抗性。在含有抗霉素的培养基中传代培养后的MA65粗制膜制剂具有正常的琥珀酸-细胞色素c氧化还原酶活性，并且在药物存在下全细胞的呼吸活性仍能持续。当在十二烷基硫酸钠（SDS）或等电聚焦凝胶上进行比较时，D98和MA65的线粒体合成蛋白相似，但通过等电聚焦检测到的一条带的标记程度存在可重复的差异。遗传分析与存在赋予抗性的细胞质定位决定因素一致。