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Chinese Science Bulletin

, Volume 47, Issue 11, pp 915–917 | Cite as

Molecular cloning, in vitro expression and enzyme activity analysis of violaxanthin de-epoxidase from Oryza sauva L.

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Abstract

The violaxanthin de-epoxidase gene was cloned from rice (Oryza sauva subsp japonica). The full length of the cDNA is 1887 bp, encoding a 446-amino acids protein with the transit peptide of 98 amino acids. The bacterial expression vector pET-Rvde was constructed and the expression quantity of the exogenous protein increased with the induction time by 0.4 mmol/L IPTG. Its molecular weight was similar with that of the native VDE. Western blotting indicated that the expressed protein has immunological reaction with the VDE polyclonal antibody. The absorbance spectrum together with xanthophyll pigments quantification by HPLC demonstrated that the expressed VDE has its enzyme activity, which can de-epoxidate violaxanthin into antheraxanthin and zeaxanthin in vitro.

Keywords

Oryza sativa L. violaxanthin de-epoxidase clone expression enzyme activity 

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References

  1. 1.
    Niyogi, K. K., Photoprotection revisited: Genetic and molecular approaches, Annu. Rev. Plant Physiol. Plant Mol. Biol., 1999, 50: 333.PubMedCrossRefGoogle Scholar
  2. 2.
    Demmig-Adams, B., Adams, W. W. III, The role of xanthophyll cycle carotenois in the protection of photosynthesis, Trends in Plant Science, 1996, 1: 21.CrossRefGoogle Scholar
  3. 3.
    Pfündel, E., Bilger, W., Regulation and possible function of violaxanthin cycle, Photosyn. Res., 1994, 42: 89.CrossRefGoogle Scholar
  4. 4.
    Eskling, M., Arvidsson, P. O., Åkerlund, H. E., The xanthophyll cycle, its regulation and components, Physiol. Plant, 1997, 100: 806.CrossRefGoogle Scholar
  5. 5.
    Müller, P., Li, X. P., Niyogi, K. K., Non-photochemical quenching: A response to excess light energy, Plant Physiol., 2001, 125: 1558.PubMedCrossRefGoogle Scholar
  6. 6.
    Lin, R. C., Ding, Z. S., Li, L. B. et al., A rapid and efficient DNA minipreparation suitable for screening transgenic plants, Plant Mol. Biol. Reptr., 2001, 19: 379a.CrossRefGoogle Scholar
  7. 7.
    Sun, Q. M., Li, L. B., Mao, D. Z. et al., High efficient expression of Lhcb2 gene from pea in E. coli and reconstitution of its expressed product with pigment in vitro, Science in China, Ser. C, 2000, 43: 464.Google Scholar
  8. 8.
    Eskling, M., Akerlund, H. E., Changes in the quantities of violaxanthin de-epoxidase, xanthophylls and ascorbate in spinach upon shift from low to high light, Photosyn. Res., 1998, 57: 41.CrossRefGoogle Scholar
  9. 9.
    Thayer, S. S., Björkman, O., Leaf xanthophyll content and composition in sun and shade determined by HPLC, Photosyn. Res., 1990, 23: 331.CrossRefGoogle Scholar
  10. 10.
    Arvidsson, P. O., Bratt, C. E., Carlsson, M. et al., Purification and identification of the violaxanthin deepoxidase as a 43kDa protein, Photosyn. Res., 1996, 49: 119.CrossRefGoogle Scholar
  11. 11.
    Bugos, R. C., Yamamoto, H. Y., Molecular cloning of violaxanthin de-epoxidase from romaine lettuce and expression in Escherichia coli, Proc. Natl. Acad. Sci. USA, 1996, 93: 6320.PubMedCrossRefGoogle Scholar
  12. 12.
    Bugos, R. C., Hieber, A. D., Yamamoto, H. Y., Xanthophyll cycle enzymes are members of the lipocalin family, the first identified from plants, J. Biol. Chem., 1998, 273: 15321.PubMedCrossRefGoogle Scholar

Copyright information

© Science in China Press 2002

Authors and Affiliations

  1. 1.Photosynthesis Research Center, Institute of BotanyChinese Academy of SciencesBeijingChina

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