DNA REPAIR GENE EXPRESSION IN MAMMALIAN CELLS

本研究提案的目标包括： 着色性干皮病DNA修复特性 成纤维细胞表达一种或多种E. coliuvr基因 (uvrA、uvrB、uvrC）稳定整合到它们的染色体中。 一 许多独立和独特的表型特性，这些 XP成纤维细胞系的克隆包括： Uvr蛋白，整合的E. coliuvr基因， 各Uvr蛋白的细胞定位，酶促 和宿主细胞通过互补受损基因的激活 显微注射Uvr蛋白和DNA存在下的存活 破坏剂。 特异性细胞核和细胞质 将检查个体修复表型的定位， 以及不同的表达浓度的影响， Uvr蛋白通过甲氨蝶呤扩增的一种共- 转染DHFR基因。 作为可行性研究的一部分，正在开发一种测定方法 用于测量人类DNA修复潜力， 评估新鲜淋巴细胞恢复免疫原性的能力。 在这种情况下，特定基因的活性被紫外线灭活 光

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.