Rosenstraus M, Chasin L A
Proc Natl Acad Sci U S A. 1975 Feb;72(2):493-7. doi: 10.1073/pnas.72.2.493.
Mutants of Chinese hamster ovary cells deficient in glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate: NADP 1-oxidoreducatse, EC 1.1.1.49) activity were isolated after mutagenesis with ethyl methane sulfonate. The mutants were induced at frequencies of about 10-4 and do not differ in growth properties from wild-type cells. They were isolated by means of a sib selection technique coupled with a histochemical stain of colonies for enzyme activity. The lack of enzyme activity is not due to a dissociable inhibitor, and is recessive in hybrid cells. Multiple mutants that lack hypoxanthine phosphoribosyltransferase activity (IMP:pyrophosphate phosphoribosyltransferase, EC 2.4.2.8) and adenine phosphoribosyltransferase activity (AMP:pyrophosphate phosphoribosyltransferase, EC 2.4.2.7) were isolated by further mutagenesis. By following segregation of wild-type phenotypes from heterozygous multiply marked hybrid cells, it was shown that the genes responsible for glucose-6-phosphate dehydrogenase activity and hypoxanthine phosphoribosyltransferase activity are linked in Chinese hamster cells, in agreement with the location of both on the X chromosome in humans. No linkage to adenosine phosphoribosyltransferase was found. The isolation of mutant cells carrying linked markers should prove useful for studying chromosomal events such as segregation, breakage, recombination, and X-chromosome reactivation.
用甲磺酸乙酯诱变后，分离出了葡萄糖-6-磷酸脱氢酶（D-葡萄糖-6-磷酸：NADP 1-氧化还原酶，EC 1.1.1.49）活性缺陷的中国仓鼠卵巢细胞突变体。这些突变体的诱导频率约为10-4，其生长特性与野生型细胞没有差异。它们是通过同胞选择技术结合对菌落进行酶活性的组织化学染色来分离的。酶活性的缺乏不是由于可解离的抑制剂，并且在杂种细胞中是隐性的。通过进一步诱变，分离出了缺乏次黄嘌呤磷酸核糖转移酶活性（IMP：焦磷酸磷酸核糖转移酶，EC 2.4.2.8）和腺嘌呤磷酸核糖转移酶活性（AMP：焦磷酸磷酸核糖转移酶，EC 2.4.2.7）的多个突变体。通过追踪杂合的多重标记杂种细胞中野生型表型的分离情况，表明负责葡萄糖-6-磷酸脱氢酶活性和次黄嘌呤磷酸核糖转移酶活性的基因在中国仓鼠细胞中是连锁的，这与它们在人类X染色体上的定位一致。未发现与腺苷磷酸核糖转移酶的连锁关系。携带连锁标记的突变细胞的分离对于研究染色体事件，如分离、断裂、重组和X染色体重新激活，应该是有用的。
