Tung P P, Respass J, Summers W C
Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, Connecticut 06520-8040, USA.
Yale J Biol Med. 1996 Nov-Dec;69(6):495-503.
A simple procedure for preparation of an affinity resin with 3'-amino thymidine linked to the carboxyl residues on 6-amino-hexanoic agarose is described. We have used this column for a rapid and simple purification of the thymidine kinase encoded by the herpes simplex virus type 1 genome. This resin has two major advantages over the most widely use used resin made with thymidine-p-nitrophenyl phosphate: first it is easily obtainable, and second, it is not subject to destruction by phosphodiesterases. The two resins are very similar in behavior and the resin made with amino thymidine has allowed us to prepare large quantities of highly purified HSV TK for crystallization studies.
本文描述了一种制备亲和树脂的简单方法,该树脂将3'-氨基胸苷连接到6-氨基己酸琼脂糖上的羧基残基上。我们使用该柱快速简单地纯化由单纯疱疹病毒1型基因组编码的胸苷激酶。与最广泛使用的由对硝基苯基磷酸胸苷制成的树脂相比,这种树脂有两个主要优点:第一,它很容易获得;第二,它不会被磷酸二酯酶破坏。这两种树脂的行为非常相似,用氨基胸苷制成的树脂使我们能够制备大量高度纯化的单纯疱疹病毒胸苷激酶用于结晶研究。