Research of Fusion Expression of Human β-Defensin-3 in Escherichia coli

Objective To achieve the fusion expression of the entire human beta-defensin-3(hBD-3) gene.Method We synthesized two oligonucleotide primers according to the codon preference of Escherichia coli.The gene was cloned into pGEX-4T-2 to establish the pGEX-4T-2-hBD-3 as the fusion expression vector by PCR.Transformed into E.coli strain DH5α,the express vector was induced and expressed by IPTG.The fusion protein GST-hBD-3 was obtained by repeated cycles of freezing and thawing,cut by thrombin to attain the recombinant hBD-3 protein.Result The result of the antibacterial peptide agarose diffusion assay shows the antibacterial activity of the rhBD-3 against the S.aureus exists,and it reaches 0.843U.Conclusion The fusion expression of the hBD-3 gene is successful.