Okumoto Kanji, Fujiki Yukio
Department of Biology, Faculty of Sciences, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka, 819-0395, Japan.
Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan.
Methods Mol Biol. 2017;1595:319-327. doi: 10.1007/978-1-4939-6937-1_29.
Cell mutants with a genetic defect affecting various cellular phenotypes are widely utilized as a powerful tool in genetic, biochemical, and cell biological research. More than a dozen complementation groups of animal somatic mutant cells defective in peroxisome biogenesis have been successfully isolated in Chinese hamster ovary (CHO) cells and used as a model system reflecting fatal human severe genetic disorders named peroxisome biogenesis disorders (PBD). Isolation and characterization of peroxisome-deficient CHO cell mutants has allowed the identification of PEX genes and the gene products peroxins, which directly leads to the accomplishment of isolation of pathogenic genes responsible for human PBDs, as well as elucidation of their functional roles in peroxisome biogenesis. Here, we describe the procedure to isolate peroxisome-deficient mammalian cell mutants from CHO cells, by making use of an effective, photo-sensitized selection method.
具有影响各种细胞表型的遗传缺陷的细胞突变体被广泛用作遗传、生化和细胞生物学研究中的强大工具。在中国仓鼠卵巢（CHO）细胞中已成功分离出十多个在过氧化物酶体生物发生方面存在缺陷的动物体细胞突变体互补群，并将其用作反映致命人类严重遗传疾病——过氧化物酶体生物发生障碍（PBD）的模型系统。过氧化物酶体缺陷型CHO细胞突变体的分离和表征使得PEX基因和过氧化物酶基因产物得以鉴定，这直接促成了导致人类PBD的致病基因的分离，以及阐明它们在过氧化物酶体生物发生中的功能作用。在此，我们描述了利用一种有效的光致敏选择方法从CHO细胞中分离过氧化物酶体缺陷型哺乳动物细胞突变体的程序。
