Aarhus University Seal / Aarhus Universitets segl

Expressivity tag: a novel tool for increased expression in Escherichia coli

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

Standard

Expressivity tag: a novel tool for increased expression in Escherichia coli. / Hansted, Jon Gade; Pietikäinen, Laura; Hög, Friederike; Sperling-Petersen, Hans Uffe; Mortensen, Kim Kusk.

I: Journal of Biotechnology, Bind 155, Nr. 3, 2011, s. 275-83.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

Harvard

Hansted, JG, Pietikäinen, L, Hög, F, Sperling-Petersen, HU & Mortensen, KK 2011, 'Expressivity tag: a novel tool for increased expression in Escherichia coli', Journal of Biotechnology, bind 155, nr. 3, s. 275-83. https://doi.org/10.1016/j.jbiotec.2011.07.013

APA

Hansted, J. G., Pietikäinen, L., Hög, F., Sperling-Petersen, H. U., & Mortensen, K. K. (2011). Expressivity tag: a novel tool for increased expression in Escherichia coli. Journal of Biotechnology, 155(3), 275-83. https://doi.org/10.1016/j.jbiotec.2011.07.013

CBE

Hansted JG, Pietikäinen L, Hög F, Sperling-Petersen HU, Mortensen KK. 2011. Expressivity tag: a novel tool for increased expression in Escherichia coli. Journal of Biotechnology. 155(3):275-83. https://doi.org/10.1016/j.jbiotec.2011.07.013

MLA

Vancouver

Hansted JG, Pietikäinen L, Hög F, Sperling-Petersen HU, Mortensen KK. Expressivity tag: a novel tool for increased expression in Escherichia coli. Journal of Biotechnology. 2011;155(3):275-83. https://doi.org/10.1016/j.jbiotec.2011.07.013

Author

Hansted, Jon Gade ; Pietikäinen, Laura ; Hög, Friederike ; Sperling-Petersen, Hans Uffe ; Mortensen, Kim Kusk. / Expressivity tag: a novel tool for increased expression in Escherichia coli. I: Journal of Biotechnology. 2011 ; Bind 155, Nr. 3. s. 275-83.

Bibtex

@article{dbf1007df73f4d1390f93da11c7c8eb2,
title = "Expressivity tag: a novel tool for increased expression in Escherichia coli",
abstract = "Protein expression in Escherichia coli is rarely trivial as low expression and insolubility are common problems. In this work we define a fusion partner, which increases expression levels similarly to the distinct function of solubility and affinity tags. This type of fusion tag we term an expressivity tag. Our work is based on earlier observations where 3' deletions of the InfB gene displays strongly increased expression levels. We have constructed progressively shortened fragments of the InfB(1-471) gene and fused gene fragments to a gfp reporter gene. A 5-fold increase in GFP expression was seen for an optimal 21 nucleotide InfB(1-21) sequence compared to gfp independently. We defined the InfB(1-21) sequence as an expressivity tag. The tag was tested for improved expression of two biotechnological important proteins streptavidin and a single chain antibody (scFv). Expression of both streptavidin and scFv(L32) was improved as evaluated by SDS-PAGE. Calculation of folding energies in the translation initiation region gave higher free energies for gfp, L32 and streptavidin when linked to InfB(1-21) than independently. InfB(1-21) did however not improve the codon usage or codon adaptation index. The expressivity tag is an important addition to the box of tools available for optimizing heterologous protein expression.",
keywords = "Animals, Biotechnology, Cloning, Molecular, Computer Simulation, Electrophoresis, Polyacrylamide Gel, Escherichia coli, Green Fluorescent Proteins, Humans, Protein Biosynthesis, Protein Folding, RNA, Messenger, Recombinant Fusion Proteins, Single-Chain Antibodies, Solubility, Spectrometry, Fluorescence, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Statistics, Nonparametric, Thermodynamics",
author = "Hansted, {Jon Gade} and Laura Pietik{\"a}inen and Friederike H{\"o}g and Sperling-Petersen, {Hans Uffe} and Mortensen, {Kim Kusk}",
note = "Copyright {\circledC} 2011 Elsevier B.V. All rights reserved.",
year = "2011",
doi = "10.1016/j.jbiotec.2011.07.013",
language = "English",
volume = "155",
pages = "275--83",
journal = "Journal of Biotechnology",
issn = "0168-1656",
publisher = "Elsevier BV",
number = "3",

}

RIS

TY - JOUR

T1 - Expressivity tag: a novel tool for increased expression in Escherichia coli

AU - Hansted, Jon Gade

AU - Pietikäinen, Laura

AU - Hög, Friederike

AU - Sperling-Petersen, Hans Uffe

AU - Mortensen, Kim Kusk

N1 - Copyright © 2011 Elsevier B.V. All rights reserved.

PY - 2011

Y1 - 2011

N2 - Protein expression in Escherichia coli is rarely trivial as low expression and insolubility are common problems. In this work we define a fusion partner, which increases expression levels similarly to the distinct function of solubility and affinity tags. This type of fusion tag we term an expressivity tag. Our work is based on earlier observations where 3' deletions of the InfB gene displays strongly increased expression levels. We have constructed progressively shortened fragments of the InfB(1-471) gene and fused gene fragments to a gfp reporter gene. A 5-fold increase in GFP expression was seen for an optimal 21 nucleotide InfB(1-21) sequence compared to gfp independently. We defined the InfB(1-21) sequence as an expressivity tag. The tag was tested for improved expression of two biotechnological important proteins streptavidin and a single chain antibody (scFv). Expression of both streptavidin and scFv(L32) was improved as evaluated by SDS-PAGE. Calculation of folding energies in the translation initiation region gave higher free energies for gfp, L32 and streptavidin when linked to InfB(1-21) than independently. InfB(1-21) did however not improve the codon usage or codon adaptation index. The expressivity tag is an important addition to the box of tools available for optimizing heterologous protein expression.

AB - Protein expression in Escherichia coli is rarely trivial as low expression and insolubility are common problems. In this work we define a fusion partner, which increases expression levels similarly to the distinct function of solubility and affinity tags. This type of fusion tag we term an expressivity tag. Our work is based on earlier observations where 3' deletions of the InfB gene displays strongly increased expression levels. We have constructed progressively shortened fragments of the InfB(1-471) gene and fused gene fragments to a gfp reporter gene. A 5-fold increase in GFP expression was seen for an optimal 21 nucleotide InfB(1-21) sequence compared to gfp independently. We defined the InfB(1-21) sequence as an expressivity tag. The tag was tested for improved expression of two biotechnological important proteins streptavidin and a single chain antibody (scFv). Expression of both streptavidin and scFv(L32) was improved as evaluated by SDS-PAGE. Calculation of folding energies in the translation initiation region gave higher free energies for gfp, L32 and streptavidin when linked to InfB(1-21) than independently. InfB(1-21) did however not improve the codon usage or codon adaptation index. The expressivity tag is an important addition to the box of tools available for optimizing heterologous protein expression.

KW - Animals

KW - Biotechnology

KW - Cloning, Molecular

KW - Computer Simulation

KW - Electrophoresis, Polyacrylamide Gel

KW - Escherichia coli

KW - Green Fluorescent Proteins

KW - Humans

KW - Protein Biosynthesis

KW - Protein Folding

KW - RNA, Messenger

KW - Recombinant Fusion Proteins

KW - Single-Chain Antibodies

KW - Solubility

KW - Spectrometry, Fluorescence

KW - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

KW - Statistics, Nonparametric

KW - Thermodynamics

U2 - 10.1016/j.jbiotec.2011.07.013

DO - 10.1016/j.jbiotec.2011.07.013

M3 - Journal article

C2 - 21801766

VL - 155

SP - 275

EP - 283

JO - Journal of Biotechnology

JF - Journal of Biotechnology

SN - 0168-1656

IS - 3

ER -