Cloning-expression Of The Antibacterial Peptide And Preliminary Study On Its Activities

Object In this study we framed the refusion gene antibacterial peptide[ Cecropin A(l ~ 8)- Magainin (1 - 12) ,CA(1 ~ 8)-MA(l ~12),CAM] vetor and expressed it in the Escherichia.coli. Thenpurified the recombinded peptide in the Intein Mediated Purificationwith an Affinity Chitin-binding Tag-IMPACT system. Finally, wetested its antibacterial activities by E.coli and S.aureus.Methods In this research, the recombinded gene was cloned into the secretion vector pTYB12 and expressed in the E.coli BL21DE3. We did the preliminary research on secretion-expression of the gene. During the purification procedure, we used the chintin beads to wash off the sundry protein, then dialysed the collecting liquid to get rid of the salinity and the other small molecular peptides. So far, we are the first ones to use the IMPACT System to purify the antibacterial peptides.Result The recombinded peptide was transformed into the E.coli BL21DE3 successfully; The recombinded antibacterial peptide 's antibacterial activities was inspected by E. coli and S.aureus , the experiment showed that recombined gene peptide has the antibacterialactivities.Conclusion The recombined gene antibacterial peptide was expressed successfully in the fusional pTYB vetor and the fusion gene peptide (CA(1 ~8)-MA(l ~12),CAM) has the antibacterial activities to E. coli and S.aurens. While how to frame a more efficient expression system should be studied further , and its other biological activities are still to be tested.