Abstract
The antimicrobial peptide CM4 is a 35-residue cationic peptide. To explore a new approach for the expression and purification of CM4 in Escherichia coli, the CM4 gene was cloned into the vector pET32a to construct an expression vector pET32a-CM4. The fusion protein Trx-CM4, purified by Ni(2+)-chelating chromatography, was cleaved by hydroxylamine hydrochloride to release recombinant CM4. Purification of recombinant CM4 was achieved by reverse HPLC chromatography, and about 1.4 mg/l active recombinant CM4 with the purity more than 98% was obtained. The recombinant CM4 showed antimicrobial activities that were similar to synthetic one.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Anti-Infective Agents / pharmacology
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Antimicrobial Cationic Peptides / biosynthesis*
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Antimicrobial Cationic Peptides / genetics*
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Antimicrobial Cationic Peptides / pharmacology
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Chromatography, Affinity
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Chromatography, High Pressure Liquid
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Escherichia coli / genetics*
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Escherichia coli / metabolism*
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Fungi / drug effects
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Genetic Vectors
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Gram-Negative Bacteria / drug effects
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Hydroxylamine / metabolism
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Microbial Sensitivity Tests
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / pharmacology
Substances
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Anti-Infective Agents
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Antimicrobial Cationic Peptides
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CM4 peptide, Bombyx mori
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Recombinant Fusion Proteins
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Hydroxylamine