Journal article
Expression and purification of SARS coronavirus proteins using SUMO-fusions
Protein expression and purification, v 42(1)
Jul 2005
PMID: 15939295
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Severe acute respiratory syndrome coronavirus (SARS-CoV) proteins belong to a large group of proteins that is difficult to express in traditional expression systems. The ability to express and purify SARS-CoV proteins in large quantities is critical for basic research and for development of pharmaceutical agents. The work reported here demonstrates: (1) fusion of SUMO (small ubiquitin-related modifier), a 100 amino acid polypeptide, to the N-termini of SARS-CoV proteins dramatically enhances expression in
Escherichia coli
cells and (2) 6× His-tagged SUMO-fusions facilitate rapid purification of the viral proteins on a large scale. We have exploited the natural chaperoning properties of SUMO to develop an expression system suitable for proteins that cannot be expressed by traditional methodologies. A unique feature of the system is the SUMO tag, which enhances expression, facilitates purification, and can be efficiently cleaved by a SUMO-specific protease to generate native protein with a desired N-terminus. We have purified various SARS-CoV proteins under either native or denaturing conditions. These purified proteins have been used to generate highly specific polyclonal antibodies. Our study suggests that the SUMO-fusion technology will be useful for enhancing expression and purification of the viral proteins for structural and functional studies as well as for therapeutic uses.
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Details
- Title
- Expression and purification of SARS coronavirus proteins using SUMO-fusions
- Creators
- Xun Zuo - LifeSensors, Inc., 271 Great Valley Parkway, Malvern, PA 19355, USAMichael R Mattern - LifeSensors, Inc., 271 Great Valley Parkway, Malvern, PA 19355, USARobin Tan - Department of Biological Sciences, National University of Singapore, 10 Kent Ridge Crescent, Singapore 119260, SingaporeShuisen Li - Drexel University, School of BioMedical Engineering, 3141 Chestnut Street, Philadelphia, PA 19104, USAJohn Hall - LifeSensors, Inc., 271 Great Valley Parkway, Malvern, PA 19355, USADavid E Sterner - LifeSensors, Inc., 271 Great Valley Parkway, Malvern, PA 19355, USAJoshua Shoo - Department of Biological Sciences, National University of Singapore, 10 Kent Ridge Crescent, Singapore 119260, SingaporeHiep Tran - LifeSensors, Inc., 271 Great Valley Parkway, Malvern, PA 19355, USAPeter Lim - Department of Biological Sciences, National University of Singapore, 10 Kent Ridge Crescent, Singapore 119260, SingaporeStefan G Sarafianos - Center for Advanced Biotechnology and Medicine (CABM), Rutgers University, Department of Chemistry and Chemical Biology, 679 Hoes Lane, Piscataway, NJ 08854-5638, USALubna Kazi - Department of Microbiology, University of Pennsylvania School of Medicine, 203A Johnson Pavilion, 36th Street and Hamilton Walk, Philadelphia, PA 19104, USASonia Navas-Martin - Department of Microbiology, University of Pennsylvania School of Medicine, 203A Johnson Pavilion, 36th Street and Hamilton Walk, Philadelphia, PA 19104, USASusan R Weiss - Department of Microbiology, University of Pennsylvania School of Medicine, 203A Johnson Pavilion, 36th Street and Hamilton Walk, Philadelphia, PA 19104, USATauseef R Butt - LifeSensors, Inc., 271 Great Valley Parkway, Malvern, PA 19355, USA
- Publication Details
- Protein expression and purification, v 42(1)
- Publisher
- Elsevier
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Microbiology and Immunology
- Web of Science ID
- WOS:000229915500014
- Scopus ID
- 2-s2.0-20444400305
- Other Identifier
- 991014877676804721
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- Collaboration types
- Domestic collaboration
- International collaboration
- Web of Science research areas
- Biochemical Research Methods
- Biochemistry & Molecular Biology
- Biotechnology & Applied Microbiology