Eur. J. Entomol. 103 (4): 867-870, 2006 | DOI: 10.14411/eje.2006.118

Expression of the melittin gene of Apis cerana cerana (Hymenoptera: Apidae) in insect cells

Wan-Jun SHI1,2, Jia-An CHENG*,1, Chuan-Xi ZHANG*,1
1 Institute of Insect Sciences, Zhejiang University, 310029 Hangzhou, China
2 Department of Resources and Environmental Science, Shanghai Jiao Tong University, 201101 Shanghai, China ; e-mails: wjshi@sjtu.edu.cn; chxzhang@zju.edu.cn; jacheng@sun.zju.edu.cn

A fragment encoding melittin cDNA from Apis cerana cerana fused with glutathione S-transferase gene was inserted into the multiple cloning site of the pBacFastHTb to construct a recombinant donor plasmid, pBacHT-GSTAccM, which was transposed to the target bacmid in E. coli (DH10) by Tn7 transposition function. Then the recombinant baculovirus Bacmid-GSTAccM was transfected into Tn-5B1-4 cells of the cabbage looper, Trichoplusia ni, mediated by lipofectin. The expressed protein of about 34 kDa was detected by Western blotting and triple antibody sandwich ELISA, indicating that the recombinant protein is the fusion protein of GSTAccM. Thin layer scanning showed that the expression level of GSTAccM was about 7% of the total cell protein. Purified and recovered recombinant melittin of A. c. cerana showed bioactivity in activating rabbit platelets to aggregate.

Keywords: Hymenoptera, Apidae, Apis cerana cerana, Chinese honeybee, melittin, insect cells, protein expression

Received: August 23, 2005; Revised: March 3, 2006; Accepted: March 3, 2006; Published: October 2, 2006  Show citation

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SHI, W., CHENG, J., & ZHANG, C. (2006). Expression of the melittin gene of Apis cerana cerana (Hymenoptera: Apidae) in insect cells. EJE, 103(4), 867-870. doi: 10.14411/eje.2006.118
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