Wheat germ cell-free expression system for protein production.
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.
Vinarov
Dmitriy A
DA
University of Wisconsin-Madison, Madison, Wisconsin.
Newman
Carrie L Loushin
CLL
University of Wisconsin-Madison, Madison, Wisconsin.
Tyler
Ejan M
EM
University of Wisconsin-Madison, Madison, Wisconsin.
Markley
John L
JL
University of Wisconsin-Madison, Madison, Wisconsin.
Shahan
Mark N
MN
Quintessence Biosciences, Inc, Madison, Wisconsin.
D000818:Animals; D002474:Cell-Free System; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D018360:Crystallography, X-Ray; D005854:Germ Cells; D006801:Humans; D019906:Nuclear Magnetic Resonance, Biomolecular; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D014158:Transcription, Genetic; D014908:Triticum / Q000235:genetics*
D012333:RNA, Messenger; D011994:Recombinant Proteins