一种从嗜血杆菌菌株中快速纯化限制性内切酶的方法。

A rapid purification method of restriction endonucleases from Haemophilus strains.

作者信息

Kopecka H

出版信息

Biochim Biophys Acta. 1975 May 23;391(1):109-20. doi: 10.1016/0005-2744(75)90157-6.

Abstract

A simple and rapid method of purification of restriction endonucleases from different Haemophilus strains is presented. By this method highly purified and stable enzymes can be obtained. Separation of different restriction activities present in the same strain is possible. This method was so far successfully used with Haemophilus influenzae, Haemophilus parainfluenzae and Haemophilus aegyptius strains. The main advantages over previously published procedures reside in the simplication of certain purification steps (for instance the BioGel A 0.5 M filtration is replaced by a hydroxyapatite batch step), elimination of exonuclease activity by fractionation with (NH4) 2SO4, separation of different restriction activities by phosphocellulose chromatography, application of this method to various strains and high purification degree of enzymes.

摘要

本文介绍了一种从不同嗜血杆菌菌株中纯化限制性内切酶的简单快速方法。通过该方法可获得高度纯化且稳定的酶。同一菌株中存在的不同限制活性能够被分离。到目前为止，该方法已成功应用于流感嗜血杆菌、副流感嗜血杆菌和埃及嗜血杆菌菌株。与先前发表的方法相比，其主要优点在于某些纯化步骤的简化（例如用羟基磷灰石批处理步骤取代BioGel A 0.5M过滤）、通过硫酸铵分级分离消除核酸外切酶活性、通过磷酸纤维素色谱法分离不同的限制活性、该方法应用于各种菌株以及酶的高纯化程度。