Department of Chemistry & Chemical Biology, Northeastern University, Boston, MA 02115, USA.
Department of Chemistry & Chemical Biology, Northeastern University, Boston, MA 02115, USA.
Trends Biotechnol. 2019 Oct;37(10):1091-1103. doi: 10.1016/j.tibtech.2019.03.011. Epub 2019 Apr 16.
DNA polymerases are critical tools in biotechnology, enabling efficient and accurate amplification of DNA templates, yet many desired functions are not readily available in natural DNA polymerases. New or improved functions can be engineered in DNA polymerases by mutagenesis or through the creation of protein chimeras. Engineering often necessitates the development of new techniques, such as selections in water-in-oil emulsions that connect genotype to phenotype and allow more flexibility in engineering than phage display. Engineering efforts have led to DNA polymerases that can withstand extreme conditions or the presence of inhibitors, as well as polymerases with the ability to copy modified DNA templates. In this review we discuss polymerases for biotechnology that have been reported along with tools to enable further development.
DNA 聚合酶是生物技术中的关键工具，能够高效准确地扩增 DNA 模板，但许多所需的功能在天然 DNA 聚合酶中并不容易获得。可以通过突变或创建蛋白质嵌合体来为 DNA 聚合酶设计新的或改进的功能。工程设计通常需要开发新技术，例如在油包水乳液中的选择，这种选择可以将基因型与表型联系起来，并比噬菌体展示提供更多的工程设计灵活性。工程设计努力已经导致了能够耐受极端条件或抑制剂存在的 DNA 聚合酶，以及具有复制修饰 DNA 模板能力的聚合酶。在这篇综述中，我们讨论了已经报道的用于生物技术的聚合酶以及能够实现进一步发展的工具。
