Expression of Eukaryotic Proteins in Soluble Form inEscherichia coli

Zhang, Yang; Olsen, David R.; Nguyen, Kimyen; Olson, Pamela; Rhodes, Eric T.; Mascarenhas, Desmond

doi:10.1006/prep.1997.0834

Cited by 100 publications

(52 citation statements)

References 32 publications

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“…Polyclonal antibody to human caveolin 1 was purchased from Upstate Biotechnology (Lake Placid, NY). Preparation of recombinant human (rh) IGFBP-3 in glycosylated (g-IGFBP-3) and non-glycosylated (ng-IGFBP-3) forms, rhIGF-I⅐rhIGFBP-3 complex, and a polyclonal antibody to intact IGFBP-3 have been described (40,41). For this work, unless otherwise specified, rhIGFBP-3 refers to non-glycosylated IGFBP-3.…”

Section: Methodsmentioning

confidence: 99%

Insulin-like Growth Factor-independent Effects Mediated by a C-terminal Metal-binding Domain of Insulin-like Growth Factor Binding Protein-3

Singh

Charkowicz

Mascarenhas

2004

Journal of Biological Chemistry

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Section: Methodsmentioning

confidence: 99%

Insulin-like Growth Factor-independent Effects Mediated by a C-terminal Metal-binding Domain of Insulin-like Growth Factor Binding Protein-3

Singh

Charkowicz

Mascarenhas

2004

Journal of Biological Chemistry

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“…When fused to eukaryotic proteins, an inactive mutant of DsbA that lacked the periplasmic signal sequence was shown to promote the soluble expression of proteins in the E. coli cytoplasm [64]. Furthermore, a comparison of DsbA and thioredoxin fusion proteins showed that DsbA increased the solubility of fusion proteins by two-to three-fold over thioredoxin [64].…”

Section: Small Ubiquitin-related Modifier (Sumo)mentioning

confidence: 99%

Recombinant protein expression and purification: A comprehensive review of affinity tags and microbial applications

Young

Britton

Robinson

2012

Biotechnology Journal

391

267

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Protein fusion tags are indispensible tools used to improve recombinant protein expression yields, enable protein purification, and accelerate the characterization of protein structure and function. Solubility-enhancing tags, genetically engineered epitopes, and recombinant endoproteases have resulted in a versatile array of combinatorial elements that facilitate protein detection and purification in microbial hosts. In this comprehensive review, we evaluate the most frequently used solubility-enhancing and affinity tags. Furthermore, we provide summaries of well-characterized purification strategies that have been used to increase product yields and have widespread application in many areas of biotechnology including drug discovery, therapeutics, and pharmacology. This review serves as an excellent literature reference for those working on protein fusion tags.

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“…Several general methods have been 72 suggested to reduce the formation of inclusion bodies, including reducing cultivation 73 temperature and altering inducer composition and concentration [16]. Other, more 74 peroxidase-specific, methodologies have been cited, including use of specific E. coli 75 strains [17], inclusion of chaperones [18] and use of leader sequences [19]. Another 76 major obstacle in the recombinant expression of hemoproteins is the limited 77 availabilty of heme and iron within a bacterial cell.…”

Section: Recombinant Peroxidase Expression 70mentioning

confidence: 99%

Horseradish and soybean peroxidases: comparable tools for alternative niches?

Ryan

Carolan

Ó’Fágáin

2006

Trends in Biotechnology

137

102

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Horseradish and soybean peroxidases (HRP and SBP, respectively) are useful 9 biotechnological tools. HRP is often termed the classical plant heme peroxidase, and 10 although it has been studied for decades our understanding has deepened since its 11 cloning and subsequent expression, which has enabled numerous mutational and 12 protein engineering studies. SBP, however, has been neglected until recently; despite 13 offering a real alternative to HRP that actually outperforms it in terms of stability. 14 SBP is now used in numerous biotechnological applications, including biosensors. 15Review of both is timely. This article summarises and discusses the main insights into 16 the structure and mechanism of HRP, with special emphasis on HRP mutagenesis, and 17 outlines its use in a variety of applications. It also reviews current knowledge and 18 applications to date of SBP, particularly biosensors. The final paragraphs speculate on 19 the future of plant heme-based peroxidases, with probable trends outlined and 20 explored. 21 22

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Expression of Eukaryotic Proteins in Soluble Form inEscherichia coli

Cited by 100 publications

References 32 publications

Insulin-like Growth Factor-independent Effects Mediated by a C-terminal Metal-binding Domain of Insulin-like Growth Factor Binding Protein-3

Insulin-like Growth Factor-independent Effects Mediated by a C-terminal Metal-binding Domain of Insulin-like Growth Factor Binding Protein-3

Recombinant protein expression and purification: A comprehensive review of affinity tags and microbial applications

Horseradish and soybean peroxidases: comparable tools for alternative niches?

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