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High–Level Expression of Human Lipocortin I in the Fission Yeast Schizosaccharomyces pombe Using a Novel Expression Vector

Bio/Technology volume 12, pages 400–404 (1994)Cite this article

Abstract

We have developed a novel expression system that allows the fission yeast, Schizosaccharomyces pombe, to be used for the efficient overproduction of heterologous proteins. As an example of the utility of this system, human lipocortin I was expressed to 50 percent of soluble protein, and 150 mg of highly purified material was obtained from 10 grams of wet cell paste. Expression of lipocortin I was driven by the human cytomegalovirus (hCMV) promoter in a vector that also contains a neomycin resistance gene (neo) under the control of the SV40 early promoter, permitting selection for increasing copy–number with increasing concentrations of the antibiotic G418. The purified protein was equivalent to its native counterpart with respect to antigenicity and biochemical properties such as phospholipase A2 inhibition, actin binding and N–terminal acetylation. We have also used this system to produce comparable amounts of other proteins including rat arginase, rat NDP–kinase and human interleukin–6.

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Author notes

  1. Hirofumi Okada

    Present address: Department of Bioengineering, Nagaoka University of Technology, Nagaoka, Niigata, 940-21, Japan

Authors and Affiliations

  1. Research Center, Asahi Glass Co. Ltd., 1150, Hazawa-Cho, Kanagawa-Ku, Yokohama, Kanagawa, Japan, 221

    Yuko Giga-Hama, Hideki Tohda, Hirofumi Okada & Hiromichi Kumagai

  2. Department of Molecular Bioregulation, Institute of Molecular and Cellular Biology for Pharmaceutical Science, Kyoto Pharmaceutical University, Yamashina-Ku, Kyoto, 607, Japan

    M. Koji Owada

  3. First Department of Biochemistry, School of Medicine, University of Tokyo, Hongo, Tokyo, 113, Japan

    Hiroto Okayama

Authors
  1. Yuko Giga-Hama
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Correspondence to Yuko Giga-Hama.

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Giga-Hama, Y., Tohda, H., Okada, H. et al. High–Level Expression of Human Lipocortin I in the Fission Yeast Schizosaccharomyces pombe Using a Novel Expression Vector. Nat Biotechnol 12, 400–404 (1994). https://doi.org/10.1038/nbt0494-400

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