Tanabe Maiko, Ishino Yoshizumi, Nishida Hirokazu
Central Research Laboratory, Hitachi Ltd., 1-280 Higashi-koigakubo, Kokubunji, Tokyo 185-8601, Japan.
Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka-shi, Fukuoka 812-8581, Japan.
Archaea. 2015 Oct 5;2015:267570. doi: 10.1155/2015/267570. eCollection 2015.
DNA ligases are indispensable in all living cells and ubiquitous in all organs. DNA ligases are broadly utilized in molecular biology research fields, such as genetic engineering and DNA sequencing technologies. Here we review the utilization of DNA ligases in a variety of in vitro gene manipulations, developed over the past several decades. During this period, fewer protein engineering attempts for DNA ligases have been made, as compared to those for DNA polymerases. We summarize the recent progress in the elucidation of the DNA ligation mechanisms obtained from the tertiary structures solved thus far, in each step of the ligation reaction scheme. We also present some examples of engineered DNA ligases, developed from the viewpoint of their three-dimensional structures.
DNA连接酶在所有活细胞中都是不可或缺的，并且在所有器官中都普遍存在。DNA连接酶广泛应用于分子生物学研究领域，如基因工程和DNA测序技术。在此，我们回顾了过去几十年来开发的DNA连接酶在各种体外基因操作中的应用。在此期间，与DNA聚合酶相比，对DNA连接酶进行蛋白质工程改造的尝试较少。我们总结了迄今为止从已解析的三级结构中获得的DNA连接机制在连接反应方案的每个步骤中的最新进展。我们还从三维结构的角度展示了一些工程化DNA连接酶的实例。
