从λT4连接酶溶原菌中快速纯化T4 DNA连接酶。
The rapid purification of T4 DNA ligase from a lambda T4 lig lysogen.
作者信息
Tait R C, Rodriguez R L, West R W
出版信息
J Biol Chem. 1980 Feb 10;255(3):813-5.
A procedure has been developed for the rapid purification of the enzyme T4 DNA ligase. The procedure involves the induction at 42 degrees C of a lambda lysogen containing the gene for T4 DNA ligase (Murray, N.E., Bruce, S.A., and Murray, K. (1979) J. Mol. Biol. 132, 493-504), followed by purification of the ligase activity by phosphocellulose and hydroxylapatite chromatography. This results in the purification of large amounts of ligase with very high specific activity. The enzyme is free of contaminating exo- and endonuclease activities and active in the ligation of DNA fragments possessing cohesive or blunt-end termini.
已开发出一种快速纯化T4 DNA连接酶的方法。该方法包括在42℃诱导含有T4 DNA连接酶基因的λ溶原菌（默里，N.E.，布鲁斯，S.A.，和默里，K.（1979年）《分子生物学杂志》132卷，493 - 504页），随后通过磷酸纤维素和羟基磷灰石层析纯化连接酶活性。这使得能够纯化出大量具有非常高比活性的连接酶。该酶不含污染性的外切核酸酶和内切核酸酶活性，并且在连接具有黏性末端或平末端的DNA片段时具有活性。
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