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| The Soluble Prokaryotic Expressed Ribosome-inactivating Protein Curcin2 with Anti-fungal Activity from Jatropha curcas L. |
| SHAO Zi-jing, JIANG Nan, YAN Hua-li, ZHAN Cheng, XU Ying, CHEN Fang |
| Key Laboratory of Bio-resources and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, 610064, China |
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Abstract The open reading frame (ORF) encoding mature curcin2 protein, a ribosomal inactivating protein induced by several kinds of stress, was amplified from genomic DNA of Jatropha curcas by PCR method.The sequence of curcin2 gene was synthesized in pMD-19-T. The gene was cloned in pGEX-6p-1 and pMAL-c5E prokaryotic expression vectors and expressed in E.coli BL21 strain. Curcin 2 protein could be expressed under different IPTG concentration, temperature and induction time but the soluble recombinant curcin2 could only be obtained in the PMAL expression system.The MBP-curcin2 fusion protein was purified through MBP TrapTM HP column and could be recognized by curcin antibody. Its antibacterial activity was tested in vitro and a better inhibition effections were found towards Gibberelle zeae (Schw.) Petch and Sclerotinia sclerotiorum (Lib) de Bary compared with those of curcin. These results may provide valuable clues and foundation to study the functions of curcin2.
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Received: 02 April 2013
Published: 25 July 2013
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