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核糖核酸酶抑制剂在脑多核糖体分离过程中的功效。

Efficacy of RNase inhibitors during brain polysome isolation.

作者信息

Gauthier D, Ven Murthy M R

出版信息

Neurochem Res. 1987 Apr;12(4):335-9. doi: 10.1007/BF00993241.

Abstract

We have investigated the efficiency of heparin, polyvinyl sulfate and yeast RNA (as competitive RNase inhibitors), liver extract (as crude preparation of liver RNase inhibitors) and DEPC (as irreversible non-competitive inhibitor) for the preparation of rat brain polysomes. Sucrose gradient sedimentation profiles, obtained from PMS, were used to determine the optimal concentration of each inhibitor. Diethylpyrocarbonate, whatever the composition of isolation buffer, was found detrimental for brain polysomes. Most of the other inhibitors where found useless or even harmful. A slight positive effect was observed with heparin 0.75 mg/mL both for total yield and sedimentation pattern. It is concluded that the utilisation of most of the widely used RNase inhibitors is of questionnable effectiveness for brain polysome preparation.

摘要

我们研究了肝素、聚乙烯硫酸酯和酵母RNA(作为竞争性核糖核酸酶抑制剂)、肝提取物(作为肝核糖核酸酶抑制剂的粗制品)以及焦碳酸二乙酯(DEPC,作为不可逆的非竞争性抑制剂)在制备大鼠脑多核糖体中的效率。从多核糖体中获得的蔗糖梯度沉降图谱用于确定每种抑制剂的最佳浓度。无论分离缓冲液的组成如何,均发现焦碳酸二乙酯对脑多核糖体有害。发现大多数其他抑制剂无用甚至有害。观察到0.75 mg/mL的肝素对总产量和沉降模式均有轻微的积极作用。得出的结论是,对于脑多核糖体的制备,大多数广泛使用的核糖核酸酶抑制剂的有效性存在疑问。

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