Fischhaber Paula L, McDaniel Lisa D, Friedberg Errol C
Department of Pathology, University of Texas Southwestern Medical Center, Dallas, USA.
Methods Enzymol. 2006;408:355-78. doi: 10.1016/S0076-6879(06)08022-0.
This chapter discusses experimental methods and protocols for the purification and preliminary characterization of DNA polymerases that are specialized for the replicative bypass (translesion DNA synthesis) of base or other types of DNA damage that typically arrest high-fidelity DNA synthesis, with particular emphasis on DNA polymerase kappa (Polkappa from mouse cells). It also describes some of the methods employed in the evaluation of mouse strains defective in genes that encode these enzymes.
本章讨论了用于纯化和初步表征DNA聚合酶的实验方法和方案,这些DNA聚合酶专门用于碱基或其他类型DNA损伤的复制性旁路(跨损伤DNA合成),这些损伤通常会阻止高保真DNA合成,尤其着重于DNA聚合酶κ(来自小鼠细胞的Polκ)。本章还描述了一些用于评估编码这些酶的基因存在缺陷的小鼠品系所采用的方法。