Cloning, expression, and antitumor activity of recombinant protein of curcin
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 wordsJatropha curcas Esherichia coli curcin expression purification recombinant protein antitumor activity
reverse transcription-polymerase chain reaction.
Unable to display preview. Download preview PDF.
- 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
- 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.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.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.Sambrook, J., Fritsch, E.F., and Maniatis, T., Molecular Cloning (A Laboratory Manual), Cold Spring Habor: Cold Spring Harbor Lab., 1989.Google Scholar
- 9.Fang, F. and Zhou, L., Modern Medical Experiment Protocol, Beijing: Beijing Medical Univ. and Xiehe Medical Univ., 1995.Google Scholar
- 10.Du, P., Medical Experimental Virus, Beijing: Publication of People’s Surgeon, 1985.Google Scholar
- 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