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1993, Bio/Technology
We have developed a versatile Escherichia coli expression system based on the use of E. coli thioredoxin (trxA) as a gene fusion partner. The broad utility of the system is illustrated by the production of a variety of mammalian cytokines and growth factors as thioredoxin fusion proteins. Although many of these cytokines previously have been produced in E. coli as insoluble aggregates or "inclusion bodies", we show here that as thioredoxin fusions they can be made in soluble forms that are biologically active. In general we find that linkage to thioredoxin dramatically increases the solubility of heterologous proteins synthesized in the E. coli cytoplasm, and that thioredoxin fusion proteins usually accumulate to high levels. Two additional properties of E. coli thioredoxin, its ability to be specifically released from the E. coli cytoplasm by osmotic shock or freeze/thaw treatments and its intrinsic thermal stability, are retained by some fusions and provide convenient purification steps. We also find that the active-site loop of E. coli thioredoxin can be used as a general site for small peptide insertions, allowing for the high level production of soluble peptides in the E. coli cytoplasm.
Journal of Immunological Methods
Versatile E. coli thioredoxin specific monoclonal antibodies afford convenient analysis and purification of prokaryote expressed soluble fusion protein1995 •
Nucleic Acids Research
A plasmid expression system for quantitative in vivo biotinylation of thioredoxin fusion proteins in Escherichia coli1998 •
Microbial Cell Factories
Expanding the molecular toolbox for Lactococcus lactis: construction of an inducible thioredoxin gene fusion expression system2011 •
Protein Expression and Purification
Expression of Three Functional Domains of Connexin 32 as Thioredoxin Fusion Proteins inEscherichia coliand Generation of Antibodies1997 •
2009 •
We report a novel fusion tag for expressing recombinant proteins in E. coli. The fusion tag is the C-terminus part of the human GMCSF gene comprising 45 amino acids, which aid in over expression of otherwise non expressible genes. Expression of hIFN a2b with this fusion tag also escapes the requirement of rare codons for expression. This is also a first report of a small fusion tag of human origin having affinity to heparin sepharose column facilitating the purification of fusion protein.
2008 •
Applied Microbiology and Biotechnology
Expression and characterization of human proinsulin fused to thioredoxin in Escherichia coli2011 •
ANNALES BOGORIENSES
Overproduction and Purification of Soluble Recombinant Human Granulocyte Colony Stimulating Factor in <i>Escherichia coli</i> Using Thioredoxin as FusionRecombinant human Granulocyte Colony Stimulating Factor (G-CSF) has been produced in a soluble form in Escherichia coli BL21 (DE3) as a fusion protein. The open reading frame of G-CSF was synthetically constructed in previous work and was codon optimized for best expression in E. coli. In this research, the gene was fused to thioredoxin (Trx) at the N-terminal in pET32 vector. The purpose of this research was to optimize the overproduction and purification processes to obtain high yield recombinant protein in soluble form, and to characterize the Trx-G-CSF fusion protein. Overproduction was performed using IPTG induction method for 3 and 6 hours. The protein was purified by Ni-NTA affinity chromatography and separated using gradient concentration of imidazole. The purified protein was then characterized by SDS-PAGE and Western Blot analysis. Further, enterokinase was used to separate G-CSF from the fusion protein. The purified form of G-CSF was subsequently characterized using Weste...
Journal of Archaeological Method and Theory
Debunking Deterministic Narratives of Technological Development Through Experimentation: A Critical Review of the Prehistory of Tin Bronze Alloying2024 •
International Journal Of Islamic Architecture
The Impact of the Mongol Conquests on Earthen Cities in Central Asia2023 •
IAEME Publications
2D IMAGE SEMANTIC SEGMENTATION FOR SELF-DRIVING CAR USING CONVOLUTION NEURAL NETWORK2021 •
2015 •
Geografia em Questão
Sobre Os (Des)Limites Territoriais: A Produção Da Natureza Enquanto Fundamento Das Dinâmicas De Fronteira2020 •
Journal of Analytical Oncology
99mTc-Labeled Bevacizumab via HYNIC for Imaging of Melanoma2014 •
2011 •
Jurnal SAINTIKOM (Jurnal Sains Manajemen Informatika dan Komputer)/Jurnal Saintikom
Topi Peringatan Bahaya Penyandang Tunanetra Berbasis Mikrokontroller Dengan Sensor Ultrasonik Menggunakan Metode Research And Development2024 •
2024 •
2018 •