›› 2006, Vol. 49 ›› Issue (1): 16-21.

• RESEARCH PAPERS • Previous Articles     Next Articles

Expression of the melittin gene from Apis cerana cerana in baculovirus system

SHI Wan-Jun, ZHANG Chuan-Xi, CHENG Jia-An   

  1. Zhejiang University
  • Online:2006-03-03 Published:2006-02-20

Abstract:

 A cDNA fragment encoding melittin from Apis cerana cerana was obtained by restriction enzyme digestion from the recombinant plasmid pGEM-AccMT, and inserted into the multiple cloning site of the pBacHTeGFPT to construct the recombinant donor plasmid pBacHT-GFPTAccM, which was transposed to the target Bacmid in E. coli (DH10) by Tn7 transposition function. Then Bacmid-GFPTAccM recombinant genome DNA was used to transfect Tn-5B1-4 cell of the cabbage looper, Trichoplusia ni, mediated by lipofectin. The expressed protein band of about 34 kD was determined by SDS-PAGE, and the thin layer scanning showed that the expression amount of GFPT-AccM fusion protein was about 3% of the total cell protein. Western blotting and the cytopathic effect of Tn cell after infection of Bacmid-GFPTAccM at different time proved that the fusion protein GFPT-AccM  had been successfully expressed in Tn-5B1-4 cell.

 

Key words: Apis cerana cerana, melittin, expression, baculovirus system, Trichopllusia ni cell line