Expression and purification of Canis interferon α in Escherichia coli using different tags

Yang, Fang; Pan, Yida; Chen, Yazhou; Tan, Shiming; Jin, Mei; Wu, Zirong; Huang, Jing

doi:10.1016/j.pep.2015.07.007

Cited by 8 publications

(3 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2c). Our result was in agreement with that from a previous study that also used this vector (Yang et al 2015). Unlike traditional separation and purification methods, which consume substantial manpower and material resources while optimizing time and temperature for enzymatic hydrolysis, the present strategy provides functional oligopeptides production with high purity (Fig.…”

Section: Discussionsupporting

confidence: 92%

Molecular modification, expression and purification of new subtype antioxidant peptide from Pinctada fucata by recombinant Escherichia coli to improve antioxidant-activity

2018

J Food Sci Technol

View full text Add to dashboard Cite

The aim of this study was to establish a system for the efficient expression and purification of new subtype of antioxidant peptide from meat (NPFMAP), which is designed by molecular modification technology based on the sequence of purified and identified antioxidant peptide from meat (PFMAP, ---------), and to better understand the relationship between structure and antioxidant activity. Meanwhile, gene codon usage was optimized and the glutathione -transferase (GST) tag of pGEX-6P-1 was added to facilitate expression and purification NPFMAP in. The results of antioxidant activity assay in vitro showed a higher antioxidant activity in NPFMAP than that in enzymatic hydrolysis digested or chemically synthesized PFMAP. In particular, the DPPH scavenging radical activity increased by about 4.7 times after molecular modification. Structural bioanalysis indicated that new subtype antioxidant peptide had spatial conformation and good hydrophilic after modification, which was confirmed by antioxidant activity assays. Thus, the proposed method could be used to obtain NPFMAP with high antioxidant activity.

show abstract

Section: Discussionsupporting

confidence: 92%

Molecular modification, expression and purification of new subtype antioxidant peptide from Pinctada fucata by recombinant Escherichia coli to improve antioxidant-activity

2018

J Food Sci Technol

View full text Add to dashboard Cite

show abstract

“…Trx is an intracellular thermostable protein of E. coli that is highly soluble expressed in its cytoplasm25. It is one of the most frequently used fusion tags for enhancing the soluble expression of recombinant proteins in E. coli cells2526. Herein, the Trx tag is incorporated to increase the solubility and stability of the target peptides after intein-mediated cleavage, particularly for peptides that are unstable in solution and more prone to form aggregates272829.…”

Section: Resultsmentioning

confidence: 99%

Recombinant production of influenza hemagglutinin and HIV-1 GP120 antigenic peptides using a cleavable self-aggregating tag

Zhao

Xing

et al. 2016

Sci Rep

View full text Add to dashboard Cite

The increasing demand for antigenic peptides in the development of novel serologic diagnostics and epitope-based vaccines requires rapid and reliable peptide synthesis techniques. Here we investigated a method for efficient recombinant expression and purification of medium- to large-sized antigenic peptides in E. coli. Previously we devised a streamlined protein expression and purification scheme based on a cleavable self-aggregating tag (cSAT), which comprised an intein molecule and a self-aggregating peptide ELK16. In this scheme, the target proteins were fused in the C-termini with cSAT and expressed as insoluble aggregates. After intein self-cleavage, target proteins were released into the soluble fraction with high yield and reasonable purity. We demonstrated the applicability of this scheme by preparing seven model viral peptides, with lengths ranging from 32 aa to 72 aa. By adding an N-terminal thioredoxin tag, we enhanced the yield of target peptides released from the aggregates. The purified viral peptides demonstrated high antigenic activities in ELISA and were successfully applied to dissecting the antigenic regions of influenza hemagglutinin. The cSAT scheme described here allows for the rapid and low-cost preparation of multiple antigenic peptides for immunological screening of a broad range of viral antigens.

show abstract

“…To our knowledge, most functional mammalian chemokines can be produced through an Escherichia coli expression system [26]. To avoid the difficult renaturation process of inclusion bodies, a practical method to obtain a soluble protein is the fusion of the target protein with a tag, which can facilitate purification without affecting the properties of the target protein [27]. Here, by using prokaryotic expression vector pET32a, grass carp CXCL20b (approximately 7.6 kDa) was fused with a 6 × His tag (approximately 1.1 kDa) and Trx tag (approximately 16.7 kDa) at its N-terminal ( Figure 1A).…”

Section: Producing Of Recombinant Grass Carp Cxcl20bmentioning

confidence: 99%

Characterization and Antimicrobial Activity of the Teleost Chemokine CXCL20b

et al. 2020

View full text Add to dashboard Cite

Fish are a potential source of diverse organic compounds with a broad spectrum of biological activities. Many fish-derived antimicrobial peptides and proteins are key components of the fish innate immune system. They are also potential candidates for development of new antimicrobial agents. CXCL20b is a grass carp (Ctenopharyngodon idella) CXC chemokine strongly transcribed at the early stage of bacterial infections, for which the immune role had not been reported to date. In the present study, we found that CXCL20b is a cationic amphipathic protein that displays potent antimicrobial activity against both Gram-positive and Gram-negative bacteria. The results of DiOC2(3) and atomic force microscopy (AFM) assays indicated that CXCL20b could induce bacterial membrane depolarization and disruption in a short time. By performing further structure-activity studies, we found that the antimicrobial activity of CXCL20b was mainly relative to the N-terminal random coil region. The central part of this cytokine representing β-sheet region was insoluble in water and the C-terminal α-helical region did not show an antimicrobial effect. The results presented in this article support the poorly understood function of CXCL20b, which fulfills an important role in bony fish antimicrobial immunity.

show abstract

Expression and purification of Canis interferon α in Escherichia coli using different tags

Cited by 8 publications

References 33 publications

Molecular modification, expression and purification of new subtype antioxidant peptide from Pinctada fucata by recombinant Escherichia coli to improve antioxidant-activity

Molecular modification, expression and purification of new subtype antioxidant peptide from Pinctada fucata by recombinant Escherichia coli to improve antioxidant-activity

Recombinant production of influenza hemagglutinin and HIV-1 GP120 antigenic peptides using a cleavable self-aggregating tag

Characterization and Antimicrobial Activity of the Teleost Chemokine CXCL20b

Contact Info

Product

Resources

About