中国生物工程杂志

CP10A, an anionic antimicrobial peptide designed from Indolicine, has a strong resistance to Gram-positive bacterial, which usually cause clinical infections. The DNA sequence of this 13 amino acids peptide was designed according to the reported CP10A peptide sequence and synthesized by PCR amplification. The sequence was cloned into the E.coli expression vector pET32a (+) and constructed the recombinant expression vector pET32a (+)-CP10A. The recombinant plasmid was transformed into E.coli AD494 and induced with IPTG. The Expression product was detected via 15% SDS-PAGE. The fusion protein was about 50% of total cellular protein and expressed as inclusion bodies. We got the pure fusion expressed CP10A protein via degeneration, Ni-NTA affinity chromatography and refolding of the complex. The purity of the refolding protein was more than 95%. It could be of great value for providing some basis to study biological activities of CP10A. It also provides some approach to study the expression of antimicrobial peptides.