Strategies for achieving high-level expression of genes in Escherichia coli.
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TLDR
Progress in the understanding of several biological processes promises to broaden the usefulness of Escherichia coli as a tool for gene expression and the remarkable increase in the availability of fusion partners offers a wide range of tools for improved protein folding, solubility, protection from proteases, yield, and secretion into the culture medium.About:
This article is published in Microbiological Research.The article was published on 1996-09-01 and is currently open access. It has received 1156 citations till now. The article focuses on the topics: Protein degradation & Codon usage bias.read more
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Journal ArticleDOI
Recombinant protein expression in Escherichia coli: advances and challenges.
TL;DR: The different approaches for the synthesis of recombinant proteins in E. coli are reviewed and recent progress in this ever-growing field is discussed.
Journal ArticleDOI
Recombinant protein expression in Escherichia coli.
TL;DR: Recent progress in the fundamental understanding of transcription, translation, and protein folding in E. coli, together with serendipitous discoveries and the availability of improved genetic tools are making this bacterium more valuable than ever for the expression of complex eukaryotic proteins.
Journal ArticleDOI
Overview of bacterial expression systems for heterologous protein production: from molecular and biochemical fundamentals to commercial systems.
TL;DR: An overview of the most commonly used promoter systems for recombinant proteins, including Bacillus brevis, Bacillusmegaterium, Bacillussubtilis, Caulobacter crescentus, other strains, and, most importantly, Escherichia coli BL21 and E. coli K12 and their derivatives are presented.
Journal ArticleDOI
Rapid protein-folding assay using green fluorescent protein.
TL;DR: This work demonstrated that the fluorescence of Escherichia coli cells expressing GFP fusions is related to the productive folding of the upstream protein domains expressed alone, providing a simple route to improving protein folding and expression by directed evolution.
Journal ArticleDOI
Soluble expression of recombinant proteins in the cytoplasm of Escherichia coli
TL;DR: New approaches to overcome obstacles by strategies that focus on either controlled expression of target protein in an unmodified form or by applying modifications using expressivity and solubility tags are reviewed.
References
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Journal ArticleDOI
Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors
TL;DR: New Escherichia coli host strains have been constructed for the E. coli bacteriophage M13 and the high-copy-number pUC-plasmid cloning vectors and mutations introduced into these strains improve cloning of unmodified DNA and of repetitive sequences.
Book ChapterDOI
Use of T7 RNA polymerase to direct expression of cloned genes.
Journal ArticleDOI
Green fluorescent protein as a marker for gene expression
TL;DR: A complementary DNA for the Aequorea victoria green fluorescent protein produces a fluorescent product when expressed in prokaryotic or eukaryotic cells, which can be used to monitor gene expression and protein localization in living organisms.
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Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes
F W Studier,B A Moffatt +1 more
TL;DR: A gene expression system based on bacteriophage T7 RNA polymerase has been developed and high levels of accumulation suggest that the RNAs are relatively stable, perhaps in part because their great length and/or stem-and-loop structures at their 3' ends help to protect them against exonucleolytic degradation.
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Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase.
D B Smith,Kevin S. Johnson +1 more
TL;DR: Plasmid expression vectors have been constructed that direct the synthesis of foreign polypeptides in Escherichia coli as fusions with the C terminus of Sj26, a 26-kDa glutathione S-transferase (GST; EC 2.5.1.18) encoded by the parasitic helminth Schistosoma japonicum.