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Expression of the melittin gene of Apis cerana cerana in Escherichia coli

Authors :
Hai-Jun Xu
Chuan-Xi Zhang
Jiaan Cheng
Wan-Jun Shi
Source :
Protein expression and purification. 37(1)
Publication Year :
2004

Abstract

A cDNA encoding melittin in Apis cerana cerana was obtained by PCR from the recombinant plasmid and cloned into the GST fusion expression vector pGEX-4T-2 for expression of the protein. The expressed protein of about 29 kDa was detected by Western blot and triple antibody sandwich ELISA, indicating that the recombinant protein is the fusion protein of GST-AccM. The expression conditions of GST-AccM fusion protein for Escherichia coli BL21 transformant were optimized. Thin layer scanning on the SDS-PAGE profiles of GST-AccM showed that the expressed protein accumulated up to about 15.2% of total protein of bacterial cells under the optimized expression condition. Purified and recovered recombinant melittin of A. cerana cerana showed bioactivity in activating rabbit platelets to aggregate.

Details

ISSN :
10465928
Volume :
37
Issue :
1
Database :
OpenAIRE
Journal :
Protein expression and purification
Accession number :
edsair.doi.dedup.....153d1a8b685feda62670e57552a6080c