Construction of eukaryotic expression vector encoding antimicrobial peptides PR-39 and its expression in 293T cell

Objective:To construct eukaryotic plasmid expressing PR-39 and test whether the plasmid can be expressed in the 293 T cell.Methods:The core coding regions(CDS) sequence of PR-39 was amplified by PCR from pBluescriptⅡSK-PR-39.Then the PR-39 gene was inserted into the eukaryotic expression plasmid pGC-FU.The resultant recombinant plasmid was confirmed by restriction enzyme cutting and sequencing,and then was designated as pGC-FU-PR-39.The recombinant plasmid pGC-FU-PR-39 was transfected into 293 T cell by Lipofectamine 2000.Transfected efficiency was observed by fluorographic unit.Western blot was used to certify whether the purpose protein was correctly expressed in 293 T cell.Results:The CDS sequence of PR-39 was correctly amplified.The recombinant plasmid was successfully constructed and could be correctly expressed in 293 T cell.Conclusion:We successfully constructed PR-39 eukaryotic expression plasmid and observed the expression of PR-39-GFP fusion protein.It lays a foundation for further studies of PR-39.