DOI: 10.1016/s0014-5793(98)01620-2. Cell‐free production and stable‐isotope labeling of milligram quantities of proteins

Cell‐free production and stable‐isotope labeling of milligram quantities of proteins

10.1016/s0014-5793(98)01620-2

Crossref journal-article

Wiley

FEBS Letters (311)

Abstract

We have improved the productivity of an Escherichia coli cell‐free protein synthesis system. First, creatine phosphate and creatine kinase were used as the energy source regeneration system, and the other components of the reaction mixture were optimized. Second, the E. coli S30 cell extract was condensed by dialysis against a polyethylene glycol solution to increase the rate of synthesis. Third, during the protein synthesis, the reaction mixture was dialyzed against a low‐molecular‐weight substrate solution to prolong the reaction. Thus, the yield of chloramphenicol acetyltransferase was raised to 6 mg/ml of reaction mixture. Stable‐isotope labeling of a protein with 13C/15N‐labeled amino acids for NMR spectroscopy was achieved by this method.

Bibliography

Kigawa, T., Yabuki, T., Yoshida, Y., Tsutsui, M., Ito, Y., Shibata, T., & Yokoyama, S. (1999). Cellâfree production and stableâisotope labeling of milligram quantities of proteins. FEBS Letters, 442(1), 15â19. Portico.

Authors 7

Takanori Kigawa (first)
Takashi Yabuki (additional)
Yasuhiko Yoshida (additional)
Michio Tsutsui (additional)
Yutaka Ito (additional)
Takehiko Shibata (additional)
Shigeyuki Yokoyama (additional)

References 23 Referenced 438

10.1016/0076-6879(91)02017-4
10.1038/nsb0894-512
10.1007/BF00211776
10.1023/A:1008276001545
10.1126/science.3055301
10.1021/bp960052l
{'key': 'e_1_2_5_8_1', 'first-page': '14', 'volume': '56', 'author': 'Davis J.', 'year': '1996', 'journal-title': 'Promega Notes Mag.'} / Promega Notes Mag. by Davis J. (1996)
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10.1006/abio.1996.0134

Dates

Type	When
Created	23 years, 1 month ago (July 25, 2002, 3:16 p.m.)
Deposited	1 year, 11 months ago (Sept. 16, 2023, 8:29 p.m.)
Indexed	4 weeks ago (July 30, 2025, 10:08 a.m.)
Issued	26 years, 7 months ago (Jan. 8, 1999)
Published	26 years, 7 months ago (Jan. 8, 1999)
Published Online	26 years, 7 months ago (Jan. 15, 1999)
Published Print	26 years, 7 months ago (Jan. 8, 1999)

Funders 0

None

BibTeX

@article{Kigawa_1999, title={Cell‐free production and stable‐isotope labeling of milligram quantities of proteins}, volume={442}, ISSN={1873-3468}, url={http://dx.doi.org/10.1016/s0014-5793(98)01620-2}, DOI={10.1016/s0014-5793(98)01620-2}, number={1}, journal={FEBS Letters}, publisher={Wiley}, author={Kigawa, Takanori and Yabuki, Takashi and Yoshida, Yasuhiko and Tsutsui, Michio and Ito, Yutaka and Shibata, Takehiko and Yokoyama, Shigeyuki}, year={1999}, month=jan, pages={15–19} }

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