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A Novel Method for High-Level Production of TEV Protease by Superfolder GFP Tag
Joint Authors
Wu, Di
Ding, Yu
Chen, Wentao
Lu, Zhisheng
Wu, Xudong
Hu, Xiaojian
Source
Journal of Biomedicine and Biotechnology
Issue
Vol. 2009, Issue 2009 (31 Dec. 2009), pp.1-8, 8 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2010-02-23
Country of Publication
Egypt
No. of Pages
8
Main Subjects
Abstract EN
Because of its stringent sequence specificity, tobacco etch virus (TEV) protease is widely used to remove fusion tags from recombinant proteins.
Due to the poor solubility of TEV protease, many strategies have been employed to increase the expression level of this enzyme.
In our work, we introduced a novel method to produce TEV protease by using visible superfolder green fluorescent protein (sfGFP) as the fusion tag.
The soluble production and catalytic activity of six variants of sfGFP-TEV was examined, and then the best variant was selected for large-scale production.
After purified by Ni-NTA affinity chromatography and Q anion exchange chromatography, the best variant of sfGFP-TEV fusion protease was obtained with purity of over 98% and yield of over 320 mg per liter culture.
The sfGFP-TEV had a similar catalytic activity to that of the original TEV protease.
Our research showed a novel method of large-scale production of visible and functional TEV protease for structural genomics research and other applications.
American Psychological Association (APA)
Wu, Xudong& Wu, Di& Lu, Zhisheng& Chen, Wentao& Hu, Xiaojian& Ding, Yu. 2010. A Novel Method for High-Level Production of TEV Protease by Superfolder GFP Tag. Journal of Biomedicine and Biotechnology،Vol. 2009, no. 2009, pp.1-8.
https://search.emarefa.net/detail/BIM-483380
Modern Language Association (MLA)
Wu, Xudong…[et al.]. A Novel Method for High-Level Production of TEV Protease by Superfolder GFP Tag. Journal of Biomedicine and Biotechnology No. 2009 (2009), pp.1-8.
https://search.emarefa.net/detail/BIM-483380
American Medical Association (AMA)
Wu, Xudong& Wu, Di& Lu, Zhisheng& Chen, Wentao& Hu, Xiaojian& Ding, Yu. A Novel Method for High-Level Production of TEV Protease by Superfolder GFP Tag. Journal of Biomedicine and Biotechnology. 2010. Vol. 2009, no. 2009, pp.1-8.
https://search.emarefa.net/detail/BIM-483380
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-483380