Sweasy J B, Yoon M S
Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, CT 06510, USA.
Mol Gen Genet. 1995 Jul 28;248(2):217-24. doi: 10.1007/BF02190803.
We used quantitative complementation assays to characterize individual DNA polymerase beta (Pol beta) mutants for their ability to function in DNA replication and DNA repair. We also describe a screen for detecting mutator activity of DNA polymerase beta mutants. By using these bioassays, together with DNA polymerase activity gels, we characterized 15 new DNA polymerase beta mutants that display a wide spectrum of phenotypes. Most of these mutants are generally defective in their ability to synthesize DNA. However, two of our Pol beta mutants show more complex phenotypes: they are able to function in DNA repair but unable to participate in DNA replication. One of our mutants displays mutator activity in vivo. Our work provides a model to study mutant mammalian enzymes in Escherichia coli with phenotypes that are otherwise difficult to assess.
我们使用定量互补分析来表征单个DNA聚合酶β（Polβ）突变体在DNA复制和DNA修复中发挥功能的能力。我们还描述了一种用于检测DNA聚合酶β突变体诱变活性的筛选方法。通过使用这些生物测定法，结合DNA聚合酶活性凝胶，我们表征了15个表现出广泛表型的新DNA聚合酶β突变体。这些突变体中的大多数在合成DNA的能力上普遍存在缺陷。然而，我们的两个Polβ突变体表现出更复杂的表型：它们能够在DNA修复中发挥作用，但不能参与DNA复制。我们的一个突变体在体内表现出诱变活性。我们的工作提供了一个模型，用于在大肠杆菌中研究具有难以评估的表型的突变哺乳动物酶。
