Recent advances in the production of proteins in insect and mammalian cells for structural biology

Nettleship, Joanne E.; Assenberg, René; Diprose, Jonathan M.; Rahman-Huq, Nahid; Owens, Raymond J.

doi:10.1016/j.jsb.2010.02.006

Cited by 90 publications

(73 citation statements)

References 111 publications

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“…In previous studies bacterial expression of recombinant human NGF demonstrated the feasibility of overproduction but also limitations in the use of E. coli as a host for expression of a protein with three intra-chain disulfide bonds. Although the other protein expression systems such as yeast, insect and mammalian cells usually produce functional protein; however, production is cost-and time-consuming and the yield is low (27). In addition, in vivo systems are not appropriate for the expression of toxic proteins.…”

Section: Discussionmentioning

confidence: 99%

Soluble Expression of Recombinant Nerve Growth Factor in Cytoplasm of Escherichia coli

2016

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Section: Discussionmentioning

confidence: 99%

Soluble Expression of Recombinant Nerve Growth Factor in Cytoplasm of Escherichia coli

2016

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“…Thus, recombinant sample production techniques, in particular using Escherichia coli as a prokaryotic expression host organism, have become commonplace for the production of proteins of interest in virtually every molecular biology laboratory. Many plasmid-based systems exist for overexpressing proteins in Escherichia coli each with their own merit, several are distributed by commercial (Jarvis, 2009;Nettleship et al, 2010). Recombinant production of protein complexes with many subunits, in particular for high-resolution structural studies, has its own challenges and intricacies.…”

Section: Challenges For Sample Preparation: New Methods For Protein Cmentioning

confidence: 99%

Structural insights into transcription complexes

Berger

Blanco

Boelens

et al. 2011

Journal of Structural Biology

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a b s t r a c tControl of transcription allows the regulation of cell activity in response to external stimuli and research in the field has greatly benefited from efforts in structural biology. In this review, based on specific examples from the European SPINE2-COMPLEXES initiative, we illustrate the impact of structural proteomics on our understanding of the molecular basis of gene expression. While most atomic structures were obtained by X-ray crystallography, the impact of solution NMR and cryo-electron microscopy is far from being negligible. Here, we summarize some highlights and illustrate the importance of specific technologies on the structural biology of protein-protein or protein/DNA transcription complexes: structure/ function analysis of components the eukaryotic basal and activated transcription machinery with focus on the TFIID and TFIIH multi-subunit complexes as well as transcription regulators such as members of the nuclear hormone receptor families. We also discuss molecular aspects of promoter recognition and epigenetic control of gene expression.

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“…Heterologous expression has been instrumental to advance protein research in the life sciences, and E. coli has largely dominated the field of recombinant expression, a trend which remains unbroken to date, although mammalian and insect cell based eukaryotic expression systems have been making their impressive mark more recently (6)(7)(8)(9). E. coli remains unsurpassed as a simple and cost effective expression host for protein production, and a very large number of plasmids and host strains are conveniently available to the research community for recombinant expression in this prokaryotic system.…”

Section: Introductionmentioning

confidence: 99%

Multiprotein Complex Production in E. coli: The SecYEG-SecDFYajC-YidC Holotranslocon

Berger

Jiang

Schulze

et al. 2017

Methods in Molecular Biology

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General rightsThis document is made available in accordance with publisher policies. Please cite only the published version using the reference above. Full terms of use are available: http://www.bristol.ac.uk/pure/about/ebr-terms SummaryA modular approach for balanced overexpression of recombinant multiprotein complexes in E. coli is described, with the prokaryotic protein secretase/insertase complex, the SecYEGSecDFYajC-YidC holotranslocon (HTL) used as an example. This procedure has been implemented here in the ACEMBL system. The protocol details the design principles of the mono-or polycistronic DNA constructs, the expression and purification of functional HTL and its association with translating ribosome nascent chain (RNC) complexes into a RNC-HTL supercomplex.1.

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Recent advances in the production of proteins in insect and mammalian cells for structural biology

Cited by 90 publications

References 111 publications

Soluble Expression of Recombinant Nerve Growth Factor in Cytoplasm of Escherichia coli

Soluble Expression of Recombinant Nerve Growth Factor in Cytoplasm of Escherichia coli

Structural insights into transcription complexes

Multiprotein Complex Production in E. coli: The SecYEG-SecDFYajC-YidC Holotranslocon

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