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A time- and cost-efficient system for high-level protein production in mammalian cells
Open accessAbstract
Most proteins for structural biology studies are produced by high-level expression in Escherichia coli. However, prokaryotic based expression systems fail to generate correctly folded functional forms of many proteins and hence a variety of eukaryotic based expression systems have been developed. Of these, yeast and baculovirus-infected insect cells currently represent the expression systems of choice for structural biologists. Here, protocols for a simple, fast and affordable method for transient protein expression in mammalian cells are reported. The results demonstrate that it combines several features necessary for the production of suitable samples for structural biology, in particular protein crystallography, namely high protein yield, straightforward purification, selenomethionine incorporation and control of N-linked glycosylation. The system is suitable for use in conventional laboratories or can be implemented in a medium- or high-throughput pipeline.
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Cited by
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Protein production and purification
Structural Genomics Consortium, Biologiques, A. E. F. D. M., Berkeley Structural Genomics Center, China Structural Genomics Consortium, Integrated Center for Structure and Function Innovation, Israel Structural Proteomics Center, Joint Center for Structural Genomics, Midwest Center for Structural Genomics, New York Structural GenomiX Research Center for Structural Genomics, Northeast Structural Genomics Consortium, et al. (2008) · Nature Methods · doi:10.1038/nmeth.f.202 · Cited by 916
Structure of RSV Fusion Glycoprotein Trimer Bound to a Prefusion-Specific Neutralizing Antibody
McLellan, J. S., Chen, M., Leung, S., Graepel, K. W., Du, X., Yang, Y., Zhou, T., Baxa, U., Yasuda, E., Beaumont, T., et al. (2013) · Science · doi:10.1126/science.1234914 · Cited by 883
Crystal structure of a human GABAA receptor
Miller, P. S., & Aricescu, A. R. (2014) · Nature · doi:10.1038/nature13293 · Cited by 710
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Metadata
- Authors
- Aricescu, A. R., Lu, W., & Jones, E. Y.
- Venue
- Acta Crystallographica Section D Biological Crystallography
- Published
- 2006-09-19
- Volume
- 62
- Issue
- 10
- Pages
- 1243-1250
- ISSN
- 0907-4449 (ISSN-L)
- OpenAlex
- W2093238184
- Cited by
- 762
- References
- 0
- FWCI
- 4.79
- Type
- journal article
- Language
- en
- Open access
- hybrid
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- Aricescu, A. R., Lu, W., & Jones, E. Y. (2006). A time- and cost-efficient system for high-level protein production in mammalian cells. Acta Crystallographica Section D Biological Crystallography. https://doi.org/10.1107/s0907444906029799
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