A novel assay for DNA ligase.
A novel assay for DNA ligase.
复制标题
一种新的 DNA 连接酶检测方法。
DOI：
10.1093/nar/19.13.3745
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发表时间：
1991
影响因子：
14.9
通讯作者：
Vishwanatha,JK
中科院分区：
文献类型：
--
作者：
Goodchild,J;Vishwanatha,JK
Replication of the lagging strand in DNA biosynthesis is discontinuous and requires a ligase activity to join together the Okazaki fragments (1). A ligase enzyme may therefore be part of the multiprotein complex involved in DNA synthesis. Here we report a new assay for DNA ligase that is more direct and simple than the usual methods (2, 3). The new method was used to search for ligase that co-fractionates with the multiprotein DNA polymerase a-primase complex from HeLa cells (4). DNA ligase was detected by its ability to join together two synthetic deoxyoligonucleotides in the presence of their complement. Polynucleotide kinase was used to 5'-phosphorylate just one of the oligonucleotides using a 32P-label.
