Desai U J, Pfaffle P K
Department of Life Sciences, Indiana State University, Terre Haute 47809, USA.
Biotechniques. 1995 Nov;19(5):780-2, 784.
The coding region of the gene for Taq DNA polymerase has been cloned into the common vector pUC18. Using a single-step procedure, large amounts of active enzyme can be purified from Escherichia coli carrying this construct. This procedure takes advantage of the thermostable properties of the DNA polymerase. This simple procedure gives very high yields of essentially homogeneous, highly active enzyme suitable for use in molecular biological applications. Yields are over two orders of magnitude greater than available with current methods.
Taq DNA聚合酶基因的编码区已被克隆到常用载体pUC18中。通过单步操作，可以从携带该构建体的大肠杆菌中纯化出大量活性酶。该方法利用了DNA聚合酶的热稳定特性。这个简单的方法能产生非常高产量的基本均匀、高活性的酶，适用于分子生物学应用。产量比现有方法高出两个数量级以上。
