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从粗糙脉孢菌野生型菌株制备无细胞翻译系统及丙酮酸激酶信使核糖核酸的翻译

Preparation of a cell-free translation system from a wild-type strain of Neurospora crassa and translation of pyruvate kinase messenger RNA.

作者信息

Devchand M, Kapoor M

出版信息

Biosci Rep. 1984 Nov;4(11):979-86. doi: 10.1007/BF01116897.

Abstract

A cell-free in vitro translation system exhibiting high activity has been developed from wild-type Neurospora crassa mycelium. The isolation is simple and fast, and the homogenization does not appear to affect the activity of mycelial proteases and nucleases. This system is capable of supporting efficient translation of exogenously added homologous RNA as demonstrated by the experiments with PK-specific mRNA. In addition, it translates heterologous RNA efficiently, shown by the translation of globin mRNA. We did not examine the Neurospora lysate for post-translational modification activity. The procedure used for the preparation of Neurospora cell-free extracts should be readily applicable to the other filamentous fungi.

摘要

已从野生型粗糙脉孢菌菌丝体开发出一种具有高活性的无细胞体外翻译系统。其分离过程简单快速,匀浆处理似乎不会影响菌丝体蛋白酶和核酸酶的活性。如针对PK特异性mRNA的实验所示,该系统能够支持外源添加的同源RNA的高效翻译。此外,如珠蛋白mRNA的翻译所示,它能有效翻译异源RNA。我们未检测粗糙脉孢菌裂解物的翻译后修饰活性。用于制备粗糙脉孢菌无细胞提取物的程序应易于应用于其他丝状真菌。

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