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Cloning and expression of rapeseed procruciferin in Escherichia coli and crystallization of the purified recombinant protein

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Abstract

Two rapeseed cruciferin cDNAs (cru2/3a and cru2/3b) were cloned and sequenced. A comparison of their DNA and protein sequences with other cruciferins, indicated cru2/3b to be a novel clone and, among them, an inherent and highly conserved sequence of twelve amino acids was identified. Procruciferin 2/3a and 2/3b were expressed in Eschericha coli, and procruciferin 2/3a was obtained in a soluble form. The expressed procruciferin 2/3a has a trimeric structure and formed crystals although the quality was not good, suggesting that this expression system is useful for protein engineering of procruciferin 2/3a.

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Authors and Affiliations

  1. Laboratory of Food Quality Design and Development, Graduate School of Agriculture, Kyoto University, Uji, Kyoto, 611-0011, Japan

    Mary Rose G. Tandang, Motoyasu Adachi & Shigeru Utsumi

Authors
  1. Mary Rose G. Tandang
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  2. Motoyasu Adachi
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  3. Shigeru Utsumi
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Tandang, M.R.G., Adachi, M. & Utsumi, S. Cloning and expression of rapeseed procruciferin in Escherichia coli and crystallization of the purified recombinant protein. Biotechnology Letters 26, 385–391 (2004). https://doi.org/10.1023/B:BILE.0000018256.90457.ee

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  • DOI: https://doi.org/10.1023/B:BILE.0000018256.90457.ee