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Transient transfection and expression of eukaryotic membrane proteins in Expi293F cells and their screening on a small scale: application for structural studies

Abstract:
Cancers, neurodegenerative and infectious diseases remain some of the leading causes of deaths worldwide. The structure-guided drug design is essential to advance drug development for these important diseases. One of the key challenges in the structure determination workflow is the production of eukaryotic membrane proteins (drug targets) of high quality. A number of expression systems have been developed for the production of eukaryotic membrane proteins. In this chapter, an optimized detailed protocol for transient transfection and expression of eukaryotic membrane proteins in Expi293F cells is presented. Testing expression and purification on a small scale allow optimizing conditions for sample preparation for downstream structural (cryo-EM) elucidation.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1007/978-1-0716-1406-8_5

Authors

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Institution:
University of Oxford
Division:
MSD
Department:
NDM
Sub department:
Human Genetics Wt Centre
Role:
Author


Publisher:
Springer Nature
Journal:
Structural Proteomics More from this journal
Volume:
2305
Pages:
105-128
Series:
Methods in Molecular Biology
Publication date:
2021-05-06
DOI:
EISSN:
1940-6029
ISSN:
1064-3745
Pmid:
33950386


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