Kaufmann W K
Department of Pathology, University of North Carolina, Chapel Hill 27599-7295.
Mutagenesis. 1988 Sep;3(5):373-80. doi: 10.1093/mutage/3.5.373.
In vitro complementation is a powerful strategy for isolation and characterization of individual components of multienzyme biochemical pathways. Application of the method to elucidate DNA metabolic pathways in prokaryotes enabled the successful identification of pathways of DNA replication and repair. In practice the technique requires the availability of genetic mutants that display defective operation of a selected biochemical pathway, and an in vitro assay system that allows detection of effective operation of the pathway. Xeroderma pigmentosum is a human disease syndrome characterized by partial or severe deficit in the operations of the nucleotidyl DNA excision repair pathway. This pathway of DNA repair appears to respond to DNA lesions which produce substantial distortion of helical structure, the best characterized of which are the UV radiation-induced pyrimidine dimers. This review summarizes a variety of approaches to analysis of the reparative deficiencies in xeroderma pigmentosum by in vitro complementation.
体外互补是分离和鉴定多酶生化途径中各个组分的一种强大策略。将该方法应用于阐明原核生物中的DNA代谢途径，成功鉴定出了DNA复制和修复途径。实际上，该技术需要有显示所选生化途径操作缺陷的遗传突变体，以及能够检测该途径有效操作的体外检测系统。着色性干皮病是一种人类疾病综合征，其特征是核苷酸DNA切除修复途径的操作部分或严重缺陷。这种DNA修复途径似乎对产生螺旋结构严重扭曲的DNA损伤有反应，其中最典型的是紫外线辐射诱导的嘧啶二聚体。本综述总结了通过体外互补分析着色性干皮病修复缺陷的多种方法。
