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使用胰蛋白酶快速高效地纯化鼠肉瘤病毒和白血病病毒。

Use of trypsin for rapid and efficient purification of murine sarcoma and leukemia virus.

作者信息

Rucheton M, Blaas D, Jeanteur P

出版信息

Biochimie. 1978;60(11-12):1333-7. doi: 10.1016/s0300-9084(79)80452-6.

Abstract

A method is described for purification of MSV-MuLV from culture supernatant of chronically infected 78A,1 rat embryo cell line. This method involves direct polyethylene glycol-NaCl precipitation of the low speed supernatant of culture fluid followed by digestion of the pellet with trypsin. This procedure efficiently disrupts large aggregates which normally entrap most of the virus. Highly purified virus can be obtained in very good yield by a combination of sedimentation velocity and isopycnic centrifugation : yields up to 100 A280 units (17 mg of protein) of purified virus per liter of culture fluid can be observed. This procedure appears well suited for large scale isolation of virion associated enzymatic activities.

摘要

本文描述了一种从长期感染的78A1大鼠胚胎细胞系的培养上清液中纯化MSV-MuLV的方法。该方法包括用聚乙二醇-氯化钠直接沉淀培养液的低速上清液,然后用胰蛋白酶消化沉淀。该程序有效地破坏了通常会截留大部分病毒的大聚集体。通过沉降速度和等密度离心相结合,可以以非常高的产率获得高度纯化的病毒:每升培养液可观察到高达100个A280单位(17毫克蛋白质)的纯化病毒产量。该程序似乎非常适合大规模分离与病毒粒子相关的酶活性。

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