Karentz D, Cleaver J E
Mol Cell Biol. 1986 Oct;6(10):3428-32. doi: 10.1128/mcb.6.10.3428-3432.1986.
Xeroderma pigmentosum (XP) is an autosomal recessive human disease, characterized by an extreme sensitivity to sunlight, caused by the inability of cells to repair UV light-induced damage to DNA. Cell fusion was used to transfer fragments of Chinese hamster ovary (CHO) chromosomes into XP cells. The hybrid cells exhibited UV resistance and DNA repair characteristics comparable to those expressed by CHO cells, and their DNA had greater homology with CHO DNA than did the DNA from XP cells. Control experiments consisted of fusion of irradiated and unirradiated XP cells and repeated exposure of unfused XP cells to UV doses used for hybrid selection. These treatments did not result in an increase in UV resistance, repair capability, or homology with CHO DNA. The hybrid cell lines do not, therefore, appear to be XP revertants. The establishment of these stable hybrid cell lines is an initial step toward identifying and cloning CHO DNA repair genes that complement the XP defect in human cells. The method should also be applicable to cloning genes for other diseases, such as ataxia-telangiectasia and Fanconi's anemia.
着色性干皮病(XP)是一种常染色体隐性人类疾病,其特征是对阳光极度敏感,这是由于细胞无法修复紫外线诱导的DNA损伤所致。细胞融合被用于将中国仓鼠卵巢(CHO)染色体片段转移到XP细胞中。杂交细胞表现出与CHO细胞相当的紫外线抗性和DNA修复特性,并且它们的DNA与CHO DNA的同源性比XP细胞的DNA更高。对照实验包括照射过的和未照射过的XP细胞的融合,以及未融合的XP细胞反复暴露于用于杂交筛选的紫外线剂量下。这些处理并未导致紫外线抗性、修复能力或与CHO DNA的同源性增加。因此,杂交细胞系似乎不是XP回复体。这些稳定杂交细胞系的建立是朝着鉴定和克隆能够弥补人类细胞中XP缺陷的CHO DNA修复基因迈出的第一步。该方法也应该适用于克隆其他疾病的基因,如共济失调毛细血管扩张症和范可尼贫血。