Expression And Research Of Antibacterial Peptide Dybowskin-1ST From The Skin Of Rana Dybowskii

The aim of this study is to express Rana dybowskii antimicrobial peptide dybowskin-1ST via Pichia Pastoris system and to analyze its antibiotic activity on six common pathogens Escherichia coli,Pseudomonas aeruginosa,Salmonella enteritidis,Staphylococcus aureus,Bacillus cereus,Klebsiella pneumoniae.The total RNA of Rana dybowskii skin was extracted by TRIzol method. The antimicrobial peptide dybowskin-1ST DNA were obtained by RT-PCR, link aim gene and double-strand His6, cloned His6-dybowskin-1ST into expression vector pPIC9K, get recombined plasmid pPIC9K-1ST. Then electrotransformed it into Pichia Pastoris GS115, generating recombinant strain expressing dybowskin-1ST. The recombinant strain was induced by the methanol and the interest protein was expressed via SDS-PAGE analysis. The interest protein was purified with Ni-Agarose and its antimicrobial activity was detected using Escherichia coli, Staphylococcus aureus, Bacillus cereus, Salmonella enteritidis, Pseudomonas aeruginosa and Klebsiella pneumoniae.The recombinant Pichia Pastoris system expressing Rana dybowskii antimicrobial peptide dybowskin-1ST was well-constructed and the peptide dybowskin-1ST showed significant antimicrobial activity on gram-positive bacteria and gram-negative bacteria. Rana dybowskii antimicrobial peptide dybowskin-1ST has a broad range of antimicrobial activity and a number of potential applications.