Stankowski L F, Hsie A W
Radiat Res. 1986 Jan;105(1):37-48.
pSV2gpt-Transformed and wild-type Chinese hamster ovary (CHO) cell lines have been used to study radiation-induced mutation at the molecular level. The transformant, designated AS52, was constructed from a hypoxanthine-guanine phosphoribosyl transferase (HPRT)-deficient CHO cell line and contains a single, functional copy of the Escherichia coli xanthine-guanine phosphoribosyl transferase (XPRT) gene (gpt) stably integrated into the Chinese hamster genome. AS52 and wild-type CHO-K1-BH4 cells exhibit similar cytotoxic responses to uv light and X rays; however, significant differences occur in mutation induction at the gpt and hprt loci. A number of HPRT and XPRT mutants which arose following irradiation were analyzed by Southern-blot hybridization. Most XPRT (21/26) and all HPRT (23/23) mutants induced by uv light exhibited hybridization patterns indistinguishable from their parental cell lines. In contrast, all XPRT (26/26) and most HPRT mutants (15/21) induced by X irradiation contained deletion mutations affecting some or all of the gpt and hprt loci, respectively. These results indicate that X rays induce predominantly deletion mutations, while uv light is likely to induce point mutations at both loci.
pSV2gpt转化的和野生型中国仓鼠卵巢(CHO)细胞系已被用于在分子水平上研究辐射诱导的突变。转化体AS52由次黄嘌呤 - 鸟嘌呤磷酸核糖转移酶(HPRT)缺陷的CHO细胞系构建而成,并且含有单个功能性大肠杆菌黄嘌呤 - 鸟嘌呤磷酸核糖转移酶(XPRT)基因(gpt)稳定整合到中国仓鼠基因组中。AS52和野生型CHO - K1 - BH4细胞对紫外线和X射线表现出相似的细胞毒性反应;然而,在gpt和hprt位点的突变诱导方面存在显著差异。通过Southern杂交分析了一些辐照后产生的HPRT和XPRT突变体。紫外线诱导的大多数XPRT(21/26)和所有HPRT(23/23)突变体表现出与其亲本细胞系无法区分的杂交模式。相比之下,X射线诱导的所有XPRT(26/26)和大多数HPRT突变体(15/21)分别包含影响部分或全部gpt和hprt位点的缺失突变。这些结果表明,X射线主要诱导缺失突变,而紫外线可能在两个位点诱导点突变。