Distance between RBS and AUG plays an important role in overexpression of recombinant proteins

被引：11

作者：

Berwal, Sunil K. ^{[1
]}

Sreejith, R. K. ^{[1
]}

Pal, Jayanta K. ^{[1
]}

机构：

[1] Univ Pune, Dept Biotechnol, Cell & Mol Biol Lab, Pune 411007, Maharashtra, India

来源：

ANALYTICAL BIOCHEMISTRY | 2010年 / 405卷 / 02期

关键词：

Ribosome binding site; Shine-Dalgarno sequence; Overexpression; AUG; Spacer; Prokaryotic expression vectors; ESCHERICHIA-COLI; INITIATION;

D O I：

10.1016/j.ab.2010.06.003

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The spacing between ribosome binding site (RBS) and AUG is crucial for efficient overexpression of genes when cloned in prokaryotic expression vectors. We undertook a brief study on the overexpression of genes cloned in Escherichia coli expression vectors, wherein the spacing between the RBS and the start codon was varied. SDS-PAGE and Western blot analysis indicated a high level of protein expression only in constructs where the spacing between RBS and AUG was similar to 40 nucleotides or more, despite the synthesis of the transcripts in the representative cases investigated. (C) 2010 Elsevier Inc. All rights reserved.

引用

页码：275 / 277

页数：3

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