Recombinant expression optimization of Human β-Defensin 3

AIM: To obtain an active recombinant human β-defensin 3(rHBD3) and a liberal supply of this product by means of optimized gene engineering technique.METHODS:Constructed expression vector pET-32a/HBD3 was transformed into Escherichia coli to obtain recombinant protein.Furthermore,the expression condition was screened and optimized.After protein purification and refolding,the product was verified in antimicrobial experiments.RESULTS:The target protein,a fusion protein with a molecular weight of about 26 kDa,was successfully expressed after IPTG induction as assessed by SDS-PAGE.The maximal expression was obtained when the expression condition was 35 ℃,with 6 hours of 0.6mmol/L IPTG induction.After purification,the antimicrobial bioactivity assay of recombinant rhBD3 showed that both the fusion protein and the mature peptide exhibited antimicrobial activity against S.aureus and E.coli.CONCLUSION:By means of gene engineering technique,massive rHBD3 with antimicrobial activities can be produced.