O'Neill J P, Brimer P A, Machanoff R, Hirsch G P, Hsie A W
Mutat Res. 1977 Oct;45(1):91-101. doi: 10.1016/0027-5107(77)90047-1.
An assay is described for the measurement of mutation induction at the hypoxanthine-guanine phosphoribosyl transferase (HGPRT) locus in Chinese hamster ovary (CHO) cells utilizing resistance to 6-thioguanine (TG). Optimal selection conditions are defined for such parameters as phenotypic expression time prior to selection, and TG concentration and cell density which permits maximum mutant recovery. The nature of the TG-resistant mutants is characterized by several physiological and biochemical methods. The data demonstrate that more than 98% of the mutant clones isolated by this selection procedure contain altered HGPRTase activity. The CHO/HGPRT system thus shows the specificity necessary for a specific gene locus mutational assay.
本文描述了一种利用对6-硫鸟嘌呤（TG）的抗性来测量中国仓鼠卵巢（CHO）细胞次黄嘌呤-鸟嘌呤磷酸核糖转移酶（HGPRT）位点突变诱导的检测方法。确定了最佳选择条件，包括选择前的表型表达时间、TG浓度和细胞密度等参数，这些参数可实现最大程度的突变体回收。通过多种生理和生化方法对TG抗性突变体的性质进行了表征。数据表明，通过该选择程序分离出的突变克隆中，超过98%含有改变的HGPRTase活性。因此，CHO/HGPRT系统显示出特定基因位点突变检测所必需的特异性。
