ArticleCurrent protocols in protein science2013
High-throughput cloning and expression of integral membrane proteins in Escherichia coli.
Article in Current protocols in protein science, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 28 citations in OpenAlex.
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- Structural insights into terminal arabinosylation of mycobacterial cell wall arabinan.Nature communications · 2025Article
- Deuterium spin relaxation of fractionally deuterated ribonuclease H using paired 475 and 950 MHz NMR spectrometers.Journal of biomolecular NMR · 2024Article
- Structural basis of peptidoglycan synthesis by E. coli RodA-PBP2 complex.Nature communications · 2023Article
- High-throughput cell-free screening of eukaryotic membrane protein expression in lipidic mimetics.Protein science : a publication of the Protein Society · 2022Article
- Structure and Function of the Bestrophin family of calcium-activated chloride channels.Channels (Austin, Tex.) · 2021Review
- Fine Sampling of Sequence Space for Membrane Protein Structural Biology.Journal of molecular biology · 2021Article
- Cryo-EM structure of arabinosyltransferase EmbB from Mycobacterium smegmatis.Nature communications · 2020Article
- Cryo-EM Structures and Regulation of Arabinofuranosyltransferase AftD from Mycobacteria.Molecular cell · 2020Article
- Subcellular localization of the five members of the human steroid 5α-reductase family.Biochimie open · 2017Article
- Atomic-level analysis of membrane-protein structure.Nature structural & molecular biology · 2016Review
- Structure of the polyisoprenyl-phosphate glycosyltransferase GtrB and insights into the mechanism of catalysis.Nature communications · 2016Article
- Escherichia coli as host for membrane protein structure determination: a global analysis.Scientific reports · 2015Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
Recently, several structural genomics centers have been established and a remarkable number of three-dimensional structures of soluble proteins have been solved. For membrane proteins, the number of structures solved has been significantly trailing those for their soluble counterparts, not least because over-expression and purification of membrane proteins is a much more arduous process. By using high-throughput technologies, a large number of membrane protein targets can be screened simultaneously and a greater number of expression and purification conditions can be employed, leading to a higher probability of successfully determining the structure of membrane proteins. This unit describes the cloning, expression, and screening of membrane proteins using high-throughput methodologies developed in the laboratory. Basic Protocol 1 describes cloning of inserts into expression vectors by ligation-independent cloning. Basic Protocol 2 describes the expression and purification of the target proteins on a miniscale. Lastly, for the targets that do express on the miniscale, Basic Protocols 3 and 4 outline the methods employed for the expression and purification of targets on a midi-scale, as well as a procedure for detergent screening and identification of detergent(s) in which the target protein is stable.
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Registered trials
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