Lohman Gregory J S, Tabor Stanley, Nichols Nicole M
New England Biolabs, Ipswich, Massachusetts, USA.
Curr Protoc Mol Biol. 2011 Apr;Chapter 3:Unit3.14. doi: 10.1002/0471142727.mb0314s94.
The DNA ligase enzyme family catalyzes the formation of a phosphodiester bond between juxtaposed 5'-phosphate and 3'-hydroxyl termini in duplex DNA. This activity can seal nicks in duplex DNA or join double-stranded DNA fragments having either blunt or cohesive ends. DNA ligases are central enzymes in molecular biology, nucleic acid research, and in next-generation sequencing applications. Reaction conditions and applications for T4 DNA ligase, E. coli DNA ligase, and thermostable DNA ligases are described in this unit. These enzymes differ in their cofactor requirements, substrate specificity, and thermal stability.
DNA连接酶家族催化双链DNA中相邻的5'-磷酸和3'-羟基末端之间形成磷酸二酯键。这种活性可以封闭双链DNA中的切口，或连接具有平端或粘性末端的双链DNA片段。DNA连接酶是分子生物学、核酸研究以及下一代测序应用中的核心酶。本单元介绍了T4 DNA连接酶、大肠杆菌DNA连接酶和热稳定DNA连接酶的反应条件及应用。这些酶在辅因子需求、底物特异性和热稳定性方面存在差异。
