Related ArticlesRelationship between recombinant protein expression and host metabolome as determined by two-dimensional NMR spectroscopy.
PLoS One. 2017;12(5):e0177233
Authors: Chae YK, Kim SH, Markley JL
Abstract
Escherichia coli has been the most widely used host to produce large amounts of heterologous proteins. However, given an input plasmid DNA, E. coli may produce soluble protein, produce only inclusion bodies, or yield little or no protein at all. Many efforts have been made to surmount these problems, but most of them have involved time-consuming and labor-intensive trial-and-error. We hypothesized that different metabolomic fingerprints might be associated with different protein production outcomes. If so, then it might be possible to change the expression pattern by manipulating the metabolite environment. As a first step in testing this hypothesis, we probed a subset of the intracellular metabolites by partially labeling it with 13C-glucose. We tested 71 genes and identified 17 metabolites by employing the two-dimensional NMR spectroscopy. The statistical analysis showed that there existed the metabolite compositions favoring protein production. We hope that this work would help devise a systematic and predictive approach to the recombinant protein production.
[NMR images] Moving from Recombinant Protein Expression to Biomolecular NMR
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Moving from Recombinant Protein Expression to Biomolecular NMR
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[NMR paper] Tracing Metabolite Footsteps of Escherichia coli Along the Time Course of Recombinant Protein Expression by Two-Dimensional NMR Spectroscopy.
Tracing Metabolite Footsteps of Escherichia coli Along the Time Course of Recombinant Protein Expression by Two-Dimensional NMR Spectroscopy.
Related Articles Tracing Metabolite Footsteps of Escherichia coli Along the Time Course of Recombinant Protein Expression by Two-Dimensional NMR Spectroscopy.
Bull Korean Chem Soc. 2012 Dec 20;33(12):4041-4046
Authors: Chae YK, Kim SH, Ellinger JJ, Markley JL
Abstract
The recombinant expression of proteins has been the method of choice to meet the demands from proteomics and structural genomics...
Chemistry and Structureof a Host–Guest Relationship:The Power of NMR and X-ray Diffraction in Tandem
Chemistry and Structureof a Host–Guest Relationship:The Power of NMR and X-ray Diffraction in Tandem
Qi-Qiang Wang, Victor W. Day and Kristin Bowman-James
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja3096762/aop/images/medium/ja-2012-096762_0013.gif
Journal of the American Chemical Society
DOI: 10.1021/ja3096762
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/jacsat/~4/Ze2zN3_VFiU
nmrlearner
Journal club
0
12-20-2012 04:48 PM
[NMR paper] Dictyostelium discoideum as expression host: isotopic labeling of a recombinant glyco
Dictyostelium discoideum as expression host: isotopic labeling of a recombinant glycoprotein for NMR studies.
Related Articles Dictyostelium discoideum as expression host: isotopic labeling of a recombinant glycoprotein for NMR studies.
Protein Expr Purif. 2000 Aug;19(3):335-42
Authors: Cubeddu L, Moss CX, Swarbrick JD, Gooley AA, Williams KL, Curmi PM, Slade MB, Mabbutt BC
The advantages of the organism Dictyostelium discoideum as an expression host for recombinant glycoproteins have been exploited for the production of an isotopically...
nmrlearner
Journal club
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11-19-2010 08:29 PM
[Nature network NMR forum] Journal club: recombinant RNA expression (0 replies)
Journal club: recombinant RNA expression (0 replies)
Recently I was thinking about my RNA project and wishing that I could express RNA in bacteria as is typically done for recombinant proteins. I looked at the July issue of Nature Methods to see a paper describing exactly the technology I was imagining. The authors created plasmids which allow the fusion of a target RNA sequence to a tRNA scaffold which is authenically processed by the cellular machinery and protected from degradation by the intrinsic stability of the tRNA. The potential benefits of this technology for NMR of RNA...
Relationship between recombinant protein expression and host metabolome as determined by two-dimensional NMR spectroscopy.
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