DNA REPAIR GENE EXPRESSION IN MAMMALIAN CELLS

这项研究提案的目标包括描述 着色性干皮病的DNA修复特性 表达一种或多种大肠杆菌UVR基因的成纤维细胞 (uvrA、uvrB、uvrC)稳定整合到它们的染色体中。一个 这些独立和独特的表型特性的数量 XP成纤维细胞系的克隆包括： UVR蛋白，整合的大肠杆菌UVR基因的结构， 各自UVR蛋白的细胞定位，酶促 和宿主细胞通过互补重新激活受损的基因 显微注射UVR蛋白与DNA存在下的存活 破坏性因素。特异核和胞浆的作用 个别修复表型的本地化将被检查为 以及改变可表达的浓度的效果 UVR蛋白通过甲氨蝶呤共扩增获得 转dhfr基因。 正在开发一种化验方法，作为可行性研究的一个组成部分。 通过一种方法测量人类的DNA修复潜力 对新鲜淋巴细胞修复的能力的评估 特定基因的活性，在这种情况下，是通过紫外线灭活的 灯。

The goals of this research proposal include the characterization of the DNA repair properties of xeroderma pigmentosum (XP) fibroblasts that express one or more of the E. coli uvr genes (uvrA, uvrB, uvrC) stably integrated into their chromosomes. A number of independent and unique phenotypic properties of these clones of XP fibroblast cell lines include: peptide maps of the Uvr proteins, structure of the integrated E. coli uvr genes, cellular localization of the respective Uvr proteins, enzymatic and host cell reactivation of damaged genes by complementing microinjected Uvr proteins and survival in the presence of DNA damaging agents. The effects of specific nuclear and cytoplasmic localization of individual repair phenotypes will be examined as well as the effects of varying the expressible concentrations of the Uvr proteins through methotrexate amplification of a co- transfected DHFR gene. An assay is being developed as a component of a feasibility study for measuring the DNA repair potential in man through an assessment of the ability of fresh lymphocytes to restore the activity of a specific gene inactivated, in this case, by ultraviolet light.