Prokaryotic expression of Ascaris antibacterial peptide Cecropin P1 gene in Escherichia coli BL21 (DE3).

Objective The complete gene sequence of Ascaris antibacterial peptide was cloned,acquired and expressed in Escherichia coli BL21(DE3) in order to obtain recombinant protein.Methods The obtained gene(Cecropin P1) sequence of the antibacterial peptide from Ascaris was modified according to E.coli preferred codons,then subcloned into the prokaryotic expression vector pET30a(+) and expressed in E.coli BL21(DE3) after the induction of IPTG.The expressed product was analyzed by SDS-PAGE.Results All products of PCR and the double restricted enzyme digestion of the two reconstructed plasmids were 113 bp in length,which was consistent with what was expected.The analysis of SDS-PAGE indicated that the reconstructed plasmid pET30a(+)-Cecropin P1 could express positively the protein with molecular weight of 3.4 ku after the induction of IPTG.Conclusion The gene plasmid pET30a(+)-Cecropin P1 was correctly constructed and the Cecropin P1 was expressed effectively in E.coli in the form of inclusion bodies,which has significance in investigating the function of the Cecropin P1 further.