Department of Dermatology, University of Michigan, Ann Arbor, MI, USA,
J Cell Commun Signal. 2008 Jun;2(1-2):25-45. doi: 10.1007/s12079-008-0026-2. Epub 2008 Sep 3.
Since molecular cloning has become routine laboratory technique, manufacturers offer countless sources of enzymes to generate and manipulate nucleic acids. Thus, selecting the appropriate enzyme for a specific task may seem difficult to the novice. This review aims at providing the readers with some cues for understanding the function and specificities of the different sources of polymerases, ligases, nucleases, phosphatases, methylases, and topoisomerases used for molecular cloning. We provide a description of the most commonly used enzymes of each group, and explain their properties and mechanism of action. By pointing out key requirements for each enzymatic activity and clarifying their limitations, we aim at guiding the reader in selecting appropriate enzymatic source and optimal experimental conditions for molecular cloning experiments.
自从分子克隆成为常规实验室技术以来，制造商提供了无数的酶源来生成和操作核酸。因此，对于新手来说，选择合适的酶来完成特定任务可能具有一定难度。本文旨在为读者提供一些线索，帮助其了解用于分子克隆的不同来源的聚合酶、连接酶、核酸酶、磷酸酶、甲基化酶和拓扑异构酶的功能和特异性。我们对每种酶组中最常用的酶进行了描述，并解释了它们的性质和作用机制。通过指出每种酶活性的关键要求并阐明其局限性，我们旨在指导读者选择合适的酶源和用于分子克隆实验的最佳实验条件。
