PURIFICATION OF AMINOACYL-TRANSFER-RNA BY AFFINITY-CHROMATOGRAPHY ON IMMOBILIZED THERMUS-THERMOPHILUS EF-TU-CENTER-DOT-GTP

被引：38

作者：

RIBEIRO, S ^{[1
]}

NOCK, S ^{[1
]}

SPRINZL, M ^{[1
]}

机构：

[1] UNIV BAYREUTH, BIOCHEM LAB, D-95440 BAYREUTH, GERMANY

来源：

ANALYTICAL BIOCHEMISTRY | 1995年 / 228卷 / 02期

关键词：

D O I：

10.1006/abio.1995.1359

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Elongation factor Tu from Thermus thermophilus containing six histidine residues on its C-terminus, EF-Tu(CHis(6)), was used for purification of aminoacyl-tRNA isoacceptors, from aminoacylated bulk. tRNA, by affinity chromatography. Preformed aminoacyl-tRNA . EF-Tu(CHis(6)). GTP ternary complexes were immobilized on Ni2+-nitriloacetic acid agarose and the aminoacyl-tRNA was eluted at high ionic strength or with buffers containing GDP. Compared to alternative methods, the reported immobilization by C-terminal histidine residues does not lead to loss of EF-Tu's affinity For aminoacyl-tRNA The method is well suited for preparative isolation of aminoacylated tRNA isoacceptors and as an analytical tool to test tRNA composition and aminoacylation of tRNA in crude cellular extracts. (C) 1995 Academic Press, Inc.

引用

页码：330 / 335

页数：6

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[11] STUDIES ON TRANSFORMATION OF ESCHERICHIA-COLI WITH PLASMIDS [J].