Nairn R S, Humphrey R M, Adair G M
University of Texas System Cancer Center, Science Park, Research Division, Smithville 78957.
Mutat Res. 1988 Jul;208(3-4):137-41. doi: 10.1016/0165-7992(88)90049-8.
DNA-mediated gene transfer (DMGT) was performed in DNA repair-proficient and UV-hypersensitive, repair-deficient Chinese hamster ovary (CHO) cell lines using the UV-irradiated thymidine kinase gene from herpes simplex virus (HSV-TK). Transformation frequencies in repair-deficient CHO cell lines declined relative to repair-proficient cells with increasing UV damage in transfected DNA; approximately 3-fold higher UV fluence was required to inactivate 50% of irradiated HSV-TK plasmid molecules in repair-proficient cells. In cotransfection experiments performed with pairs of HSV-TK plasmids containing linker insertion mutations in TK coding sequences, moderate UV damage in plasmid DNA enhanced the yield of TK+ transformants resulting from homologous recombination between HSV-TK sequences up to 4-fold. These results suggest that UV damage in DNA can stimulate transformation of mammalian cells dependent on intermolecular DNA homology.
使用来自单纯疱疹病毒的紫外线照射的胸苷激酶基因(HSV-TK),在DNA修复功能正常和紫外线超敏、修复缺陷的中国仓鼠卵巢(CHO)细胞系中进行了DNA介导的基因转移(DMGT)。随着转染DNA中紫外线损伤的增加,修复缺陷的CHO细胞系中的转化频率相对于修复功能正常的细胞有所下降;在修复功能正常的细胞中,使50%的经照射的HSV-TK质粒分子失活所需的紫外线通量大约高3倍。在用在TK编码序列中含有接头插入突变的成对HSV-TK质粒进行的共转染实验中,质粒DNA中的中度紫外线损伤使HSV-TK序列之间同源重组产生的TK+转化体产量提高了4倍。这些结果表明,DNA中的紫外线损伤可以刺激依赖分子间DNA同源性的哺乳动物细胞转化。