Graf H
Biochim Biophys Acta. 1979 Sep 27;564(2):225-34. doi: 10.1016/0005-2787(79)90221-1.
Conditions for the ligation with T4 induced DNA ligase of two DNA molecules via their complementary sticky ends have been established which lead preferentially to the formation of hybrid molecules. This is demonstrated with two combinations of parent molecules varying greatly in their relative molecular weights. In one case the intact hybrid molecule could be directly isolated. In addition a DNA dependent quantitative electrophoretic assay for DNA ligase activity is described which does not need a radioactively labeled substrate. The ligation procedure has been shown to be useful in molecular cloning experiments.
已经确定了通过两个DNA分子的互补粘性末端用T4诱导的DNA连接酶进行连接的条件，这些条件优先导致杂交分子的形成。这通过两种相对分子量差异很大的亲本分子组合得到了证明。在一种情况下，可以直接分离完整的杂交分子。此外，还描述了一种用于DNA连接酶活性的DNA依赖性定量电泳测定法，该方法不需要放射性标记的底物。该连接程序已被证明在分子克隆实验中是有用的。
