Mayer, Michael, Dailey, Tamara A., Baucom, Clay, Supernak, Jill L, Grady, Michael C, Hawk, Harris E, Dailey, Harry A. (2004) Expression of human proteins at the Southeast Collaboratory for Structural Genomics. Journal of Structural and Functional Genomics, 5 (1-2). pp. 159-165. ISSN 1345-711X. (doi:10.1023/B:JSFG.0000029202.77832.34) (Access to this publication is currently restricted. You may be able to access a copy if URLs are provided) (KAR id:84505)
PDF
Publisher pdf
Language: English Restricted to Repository staff only |
|
|
![]() |
Official URL: http://dx.doi.org/10.1023/B:JSFG.0000029202.77832.... |
Abstract
The human protein production group at the Southeast Collaboratory for Structural Genomics is charged with producing human proteins for both X-ray crystallography and NMR structural studies. Eukaryotic, and human proteins in particular, are notoriously difficult to express in bacterial systems. For various reasons, T7-based expression often results in protein expressed in an insoluble form. Overcoming this requires either introduction of a step to screen expression conditions or inclusion of a troublesome refolding step during purification. Our laboratory uses a trc-based expression vector that addresses many of the difficulties of the commonly used T7-based expression systems. Proteins expressed under the trc promoter, a weak promoter compared to the strong T7 promoter, are produced in a soluble form and include necessary cofactors. The details of this system will be discussed.
Item Type: | Article |
---|---|
DOI/Identification number: | 10.1023/B:JSFG.0000029202.77832.34 |
Subjects: | Q Science |
Divisions: | Divisions > Division of Natural Sciences > Biosciences |
Depositing User: | Jill Shepherd |
Date Deposited: | 27 Nov 2020 18:29 UTC |
Last Modified: | 16 Nov 2021 10:27 UTC |
Resource URI: | https://kar.kent.ac.uk/id/eprint/84505 (The current URI for this page, for reference purposes) |
- Export to:
- RefWorks
- EPrints3 XML
- BibTeX
- CSV
- Depositors only (login required):