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Expression and Purification of Recombinant Mouse Interleukin-4 and -6 from Transgenic Rice Seeds

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Abstract

Transgenic rice seed can be utilized as a bioreactor to produce high-value recombinant proteins. Mouse interleukin 4 (mIL-4) and mIL-6 were specifically expressed as secretory proteins in rice endosperm by ligating the N-terminal glutelin B-1 (GluB-1) signal peptide and the C-terminal KDEL endoplasmic reticulum retention signal under control of the endosperm-specific GluB-1 promoter. In the transgenic rice seed, mIL-4 and mIL-6 accumulated in levels up to 0.43 mg/g grain and 0.16 mg/g grain, respectively. The reducing agents and detergents required for extraction from the transgenic rice seeds differed between the two proteins, indicating differences in their intracellular localization within the endosperm cell. Purified mIL-4 and mIL-6 exhibited high activity and very low endotoxin contamination.

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Acknowledgments

This work was supported by ‘Genomics and Agricultural Innovation, GMC0004’ from the Ministry of Agriculture, Forestry, and Fisheries of Japan

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Authors and Affiliations

  1. Preventec, Inc., 1-2 Owashi, Tsukuba, Ibaraki, 305-8634, Japan

    Yoshihiro Fujiwara & Kenji Sekikawa

  2. Transgenic Crop Research and Development Center, National Institute of Agrobiological Sciences, Kannondai 3-1-3, Tsukuba, Ibaraki, 305-8604, Japan

    Lijun Yang & Fumio Takaiwa

Authors
  1. Yoshihiro Fujiwara
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  2. Lijun Yang
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  3. Fumio Takaiwa
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  4. Kenji Sekikawa
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Correspondence to Yoshihiro Fujiwara.

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Fujiwara, Y., Yang, L., Takaiwa, F. et al. Expression and Purification of Recombinant Mouse Interleukin-4 and -6 from Transgenic Rice Seeds. Mol Biotechnol 58, 223–231 (2016). https://doi.org/10.1007/s12033-016-9920-7

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  • DOI: https://doi.org/10.1007/s12033-016-9920-7

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