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Please use this identifier to cite or link to this item: http://hdl.handle.net/10119/7871

Title: Stable-isotope labeling using an inducible viral infection system in suspension-cultured plant cells
Authors: Ohki, Shinya
Dohi, Koji
Tamai, Atsushi
Takeuchi, Makoto
Mori, Masashi
Keywords: Inducible virus vector
stable-isotope labeling
Issue Date: 2008-12
Publisher: Springer
Magazine name: Journal of Biomolecular NMR
Volume: 42
Number: 4
Start page: 271
End page: 277
DOI: 10.1007/s10858-008-9283-x
Abstract: We established a novel strategy for preparing uniformly stable isotope-labeled proteins by using suspension-cultured plant cells and an inducible virus vector encoding the research target. By using this new method, we demonstrated the expression of three proteins, namely, Escherichia coli dihydrofolate reductase (DHFR), chicken calmodulin (CaM), and porcine protein kinase C-dependent protein phosphatase-1 inhibitor with a molecular mass of 17-kDa (CPI-17). In addition, we successfully expressed bovine pancreatic trypsin inhibitor (BPTI), which contains three pairs of disulfide bonds, as the soluble form. In the most efficient case, as little as 50 ml culture yielded 3-4 mg ^<15>N-labeled protein suitable for NMR experiments. The ^1H-^<15>N HSQC spectra of all of these proteins clearly indicated that their structures were identical to those of their counterparts reported previously. Thus, the present results suggest that our novel protocol is a potential method for NMR sample preparation.
Rights: This is the author-created version of Springer, Shinya Ohki, Koji Dohi, Atsushi Tamai, Makoto Takeuchi and Masashi Mori, Journal of Biomolecular NMR, 42(4), 2008, 271-277. The original publication is available at www.springerlink.com, http://dx.doi.org/10.1007/s10858-008-9283-x
URI: http://hdl.handle.net/10119/7871
Material Type: author
Appears in Collections:f10-1. 雑誌掲載論文 (Journal Articles)

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