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GFP-Based expression screening of membrane proteins in insect cells using the baculovirus system
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Hu, Nien-Jen, Rada, Heather, Rahman, Nahid, Nettleship, Joanne E., Bird, Louise, Iwata, So, Drew, David, Cameron, Alexander and Owens, Raymond J. (2014) GFP-Based expression screening of membrane proteins in insect cells using the baculovirus system. In: Owens, Raymond J. , (ed.) Structural Proteomics : High-Throughput Methods. Humana Press, pp. 197-209. ISBN 9781493922291
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Official URL: http://dx.doi.org/10.1007/978-1-4939-2230-7_11
Abstract
A key step in the production of recombinant membrane proteins for structural studies is the optimization of protein yield and quality. One commonly used approach is to fuse the protein to green fluorescent protein (GFP), enabling expression to be tracked without the need to purify the protein. Combining fusion to green fluorescent protein with the baculovirus expression system provides a useful platform for both screening and production of eukaryotic membrane proteins. In this chapter we describe our protocol for the expression screening of membrane proteins in insect cells using fusion to GFP as a reporter. We use both SDS-PAGE with in-gel fluorescence imaging and fluorescence-detection size-exclusion chromatography (FSEC) to screen for expression.
Item Type: | Book Item | ||||
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Subjects: | Q Science > QP Physiology | ||||
Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) > Biological Sciences ( -2010) Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) |
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Library of Congress Subject Headings (LCSH): | Membrane proteins, Baculoviruses, Green fluorescent protein | ||||
Journal or Publication Title: | Methods in Molecular Biology (Clifton, N.J.) | ||||
Publisher: | Humana Press | ||||
ISBN: | 9781493922291 | ||||
ISSN: | 1064-3745 | ||||
Book Title: | Structural Proteomics : High-Throughput Methods | ||||
Editor: | Owens, Raymond J. | ||||
Official Date: | 22 November 2014 | ||||
Dates: |
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Volume: | 1261 | ||||
Page Range: | pp. 197-209 | ||||
DOI: | 10.1007/978-1-4939-2230-7_11 | ||||
Status: | Peer Reviewed | ||||
Publication Status: | Published | ||||
Access rights to Published version: | Restricted or Subscription Access |
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