Molecular Cloning, Expression, Purification, and Functional Characterization of Palustrin-2CE, an Antimicrobial Peptide of Rana chensinensis

作者：Sun, Yan; Li, Qian; Li, Zhi^*; Zhang, Yuan; Zhao, Jie; Wang, Lin

来源：Bioscience Biotechnology and Biochemistry, 2012, 76(1): 157-162.

DOI：10.1271/bbb.110672

• antimicrobial peptide
• Rana chensinensis
• soluble protein
• affinity purification
• antibacterial activity

Antimicrobial peptides are effector molecules of the innate immunity of amphibians. Here, one antimicrobial peptide cDNA precursor, prepropalustrin-2CE3, from the tadpole of the Chinese brown frog Rana chensinensis was cloned. The coding sequence corresponding to the mature palustrin-2CE peptide was subcloned into pGEX-6p-1. The soluble GST-palustrin-2CE fusion protein was successfully expressed in the BL21(DE3)-pLysS strain at 16 C, and the proportion of the fusion protein reached 35%-39% of the total cellular protein. After removal of the GST-fusion tag, the purity of the palustrin-2CE obtained by Sephadex G50 chromatography was about 97%. Moreover, the purified palustrin-2CE displayed obviously inhibitory activities against the sensitive bacteria Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, and Escherichia coli, and multi-drug resistant S. aureus and E. coli. These findings suggest that the tadpole of the Chinese brown frog is a unique source of antimicrobial peptides and indicates the therapeutic potential of the palustrin-2CE peptide.

• 出版日期2012-1
• 单位陕西师范大学