Wheat germ cell-free expression system for protein production

Current Protocols in Protein Science
Dmitriy A VinarovMark N Shahan

Abstract

The Center for Eukaryotic Structural Genomics, in cooperation with Ehime University and CellFree Sciences, has developed a novel wheat germ cell-free technology for the production of eukaryotic proteins. Protein production and purification are robust and scalable for high-throughput applications. The protocols have been used to express and purify proteins from Arabidopsis thaliana, human, mouse, rat and zebra fish. This unit describes expression and purification protocols for both small-scale testing (microgram) and large-scale production (milligram) of N-His6- and N-GST-tagged proteins. The methods described in this unit can be used to produce both unlabeled and labeled proteins required for structure-based determinations by NMR spectroscopy or X-ray crystallography.

References

Jan 1, 1982·Molecular & General Genetics : MGG·B HenrichR Plapp
Aug 1, 1996·European Journal of Biochemistry·D M KimS Yokoyama
Jan 19, 2000·Proceedings of the National Academy of Sciences of the United States of America·K MadinY Endo
Jun 3, 2000·Biotechnology Progress·D M Kim, J R Swartz
Dec 5, 2000·Nature Structural Biology·S YokoyamaS Kuramitsu
Nov 1, 2002·Proceedings of the National Academy of Sciences of the United States of America·Tatsuya SawasakiYaeta Endo
Jan 28, 2003·Current Opinion in Chemical Biology·Shigeyuki Yokoyama
May 23, 2003·Protein Science : a Publication of the Protein Society·Eugene Hayato MoritaToshiyuki Kohno
Nov 19, 2003·European Journal of Biochemistry·Takayasu KawasakiYaeta Endo
Jan 20, 2004·European Journal of Biochemistry·Christian KlammtFrank Bernhard
Jan 31, 2004·Journal of Biomolecular NMR·Jianxia ShiDavid E Wemmer
Mar 11, 2004·European Journal of Biochemistry·Namthip ChumpolkulwongShigeyuki Yokoyama
Mar 31, 2004·Biotechnology and Bioengineering·Gang Yin, James R Swartz
Mar 23, 2005·Nature Methods·Dmitriy A VinarovJohn L Markley

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Citations

Jan 9, 2009·Journal of Structural and Functional Genomics·John L MarkleyZsolt Zolnai
Mar 10, 2010·Journal of Structural and Functional Genomics·William H EschenfeldtAndrzej Joachimiak
Mar 10, 2011·Proteins·E Sethe BurgieGeorge N Phillips
Dec 14, 2011·Protein Science : a Publication of the Protein Society·Eva-Maria E UhlemannOleg Y Dmitriev
Aug 26, 2006·The FEBS Journal·Dmitriy A VinarovJohn L Markley
Nov 6, 2018·Nucleic Acids Research·Venkata Rajesh YellaManju Bansal
Apr 13, 2021·Frontiers in Molecular Biosciences·Marie-Laure FogeronAnja Böckmann

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