Russian Journal of Plant Physiology

, Volume 54, Issue 2, pp 202–206 | Cite as

Cloning, expression, and antitumor activity of recombinant protein of curcin

  • M. J. Luo
  • W. X. Liu
  • X. Y. Yang
  • Y. Xu
  • F. Yan
  • P. Huang
  • F. Chen
Research Papers


Curcin, a protein isolated from the seeds of Jatropha curcas can be used as a cell-killing agent. To elaborate the purification methods and investigate the antitumor activity of the recombinant protein, the fragment encoding the mature protein of curcin was inserted into E. coli strain M15 and the recombinant strain was induced to express by the optimum inducer (0.5 mM isopropyl-β-D-thiogalactopyranoside). The recombinant protein was expressed in the form of the inclusion body and was purified by Ni-NTA affinity chromatography. The protein of interest was incubated with the tumor cells at various concentrations for different time. It was shown that the target protein could inhibit the growth of NCL-H446, SGC-7901, and S180 at a very low concentration.

Key words

Jatropha curcas Esherichia coli curcin expression purification recombinant protein antitumor activity 



guanidine chloride




phosphate-buffered saline


phenylmethylsulfonyl fluoride


ribosome-inactivating protein


reverse transcription-polymerase chain reaction.


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  1. 1.
    Girbes, T., Ferreras, J.M., Arias, F.J., and Stirpe, F., Description, Distribution, Activity and Phylogenetic Relationship of Ribosome-Inactivating Proteins in Plants, Fungi and Bacteria, Mini-Reviews in Med. Chem., 2004, vol. 4, pp. 461–467.Google Scholar
  2. 2.
    Nielsen, K. and Boston, R.S., Ribosome-Inactivating Proteins: A Plant Perspective, Annu. Rev. Plant Physiol. Plant Mol. Biol., 2001, vol. 52, pp. 785–816.PubMedCrossRefGoogle Scholar
  3. 3.
    Park, S.W., Vepachedu, R., Sharma, N., and Vivanco, J.M., Ribosome-Inactivating Proteins in Plant Biology, Planta, 2004, vol. 219, pp. 1093–1096.PubMedCrossRefGoogle Scholar
  4. 4.
    Stripe, F., Ribosome-Inactivating Proteins, Toxicin, 2004, vol. 44, pp. 371–379.CrossRefGoogle Scholar
  5. 5.
    Stripe, F., Pession-Brizzi, A., and Lorenzoni, E., Studies on the Proteins from Seeds of Croton tiglium and of Jatropha curcas, Biochem. J., 1976, vol. 156, pp. 1–6.Google Scholar
  6. 6.
    Zhang, N., Wei, Z., He, J., Du, L., and Liang, H., An Efficient and Economic Method for Preparation of High Quality Plant RNA, Progr. Biochem. Biophys., 2004, vol. 31, pp. 947–950.Google Scholar
  7. 7.
    Lin, J., Chen, Y., Xu, Y., Yan, F., Tang, L., and Chen, F., Cloning and Expression of Curcin, a Ribosome-Inactivating Protein from the Seeds of Jatropha curcas, Acta Bot. Sin., 2003, vol. 45, pp. 858–863.Google Scholar
  8. 8.
    Sambrook, J., Fritsch, E.F., and Maniatis, T., Molecular Cloning (A Laboratory Manual), Cold Spring Habor: Cold Spring Harbor Lab., 1989.Google Scholar
  9. 9.
    Fang, F. and Zhou, L., Modern Medical Experiment Protocol, Beijing: Beijing Medical Univ. and Xiehe Medical Univ., 1995.Google Scholar
  10. 10.
    Du, P., Medical Experimental Virus, Beijing: Publication of People’s Surgeon, 1985.Google Scholar
  11. 11.
    Vepachedu, R., Park, S.W., Sharma, N., and Vivanco, J.M., Bacterial Expression and Enzymatic Activity Analysis of ME1, a Ribosome-Inactivating Protein from Mirabilis expansa, Protein Expres. Purif., 2005, vol. 40, pp. 142–151.CrossRefGoogle Scholar
  12. 12.
    Goto, L.S., Beltramini, L.M., de Moraes D.I., Moreira, R.A., and de Araujo, A.P., Abrus pulchellus Type-2 RIP, Pulchellin: Heterologous Expression and Refolding of the Sugar-Binding B Chain, Protein Expres. Purif., 2003, vol. 31, pp. 12–18.CrossRefGoogle Scholar
  13. 13.
    Guo, C., Li, Z., Shi, Y., Xu, M., John, G.W., Wolfgong, E.T., and Yuan, J., Intein-Mediated Fusion Expression, High Efficient Refolding, and One-Step Purification of Gelonin Toxin, Protein Expres. Purif., 2004, vol. 37, pp. 361–367.CrossRefGoogle Scholar
  14. 14.
    Rudolph, R. and Lilie, H., In Vitro Folding of Inclusion Body Proteins, FASEB J., 1996, vol. 10, pp. 49–57.PubMedGoogle Scholar
  15. 15.
    Ceciliani, F., Caramori, T., Ronchi, S., Tedeschi, G., Mortarino, M., and Galizzi, A., Cloning, Overexpression, and Purification of Escherichia coli Quinolinate Synthetase, Protein Expres. Purif., 2000, vol. 18, pp. 64–70.CrossRefGoogle Scholar
  16. 16.
    Olsnes, S., Fernandez-Puentes, C., Carrasco, L., and Vazquea, D., Ribosome Inactivation by the Toxic Lectins Abrin and Ricin Kinetics of the Enzymic Activity of the Toxin A-Chain, Eur. J. Biochem., 1975, vol. 60, pp. 281–288.PubMedCrossRefGoogle Scholar
  17. 17.
    Zamboni, M., Brigotti, M., Rambelli, F., Montanaro, L., and Sperti, S., High-Pressure-Liquid-Chromatographic and Fluorimetric Methods for the Determination of Adenine Released from Ribosomes by Ricin and Gelonin, Biochem. J., 1989, vol. 259, pp. 639–643.PubMedGoogle Scholar
  18. 18.
    Endo, Y. and Tsurugi, K., RNA N-Glycosidase Activity of Ricin A-Chain. Mechanism of Action of the Toxic Lectin Ricin on Eukaryotic Ribosomes, J. Biol. Chem., 1987, vol. 262, pp. 8128–8130.PubMedGoogle Scholar
  19. 19.
    Brigotti, M., Barbieri, L., Valbonesi, P., Stirpe, F., Montanaro, L., and Sperti, S., A Rapid and Sensitive Method to Measure the Enzymatic Activity of Ribosome-Inactivating Proteins, Nucleic Acids Res., 1998, vol. 26, pp. 4306–4307.PubMedCrossRefGoogle Scholar
  20. 20.
    Lin, J., Yan, F., Tang, L., and Chen, F., Isolation, Purification and Functional Investigation in the N-Glycosidase Activity of Curcin from the Seeds of Jatropha curcas, High-Technic Commun., 2002, vol. 11, pp. 36–40.Google Scholar
  21. 21.
    Peumans, W.J., Hao, Q., and van Damme, E.J.M., Ribosome-Inactivating Proteins from Plants: More than RNA N-Glycosidases? FASEB J., 2001, vol. 13, pp. 1493–1506.CrossRefGoogle Scholar
  22. 22.
    Narayanan, S., Surendranath, K., Bora, N., Surolia, A., and Karande, A.A., Ribosome Inactivating Proteins and Apoptosis, FEBS Lett., 2005, vol. 579, pp. 1324–1331.PubMedCrossRefGoogle Scholar
  23. 23.
    Stirpe, F. and Barbieri, L., Ribosome-Inactivating Proteins up to Date, FEBS Lett., 1986, vol. 195, pp. 1–8.PubMedCrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2007

Authors and Affiliations

  • M. J. Luo
    • 1
  • W. X. Liu
    • 2
  • X. Y. Yang
    • 1
  • Y. Xu
    • 1
  • F. Yan
    • 1
  • P. Huang
    • 2
  • F. Chen
    • 1
  1. 1.College of Life and ScienceSichuan University 24ChengduChina
  2. 2.Chengdu Institute for Family PlanningChengduChina

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