Construction And Prokaryotic Expression Of Novel Fusion Gene Of Bovine Neutrophil Beta Defensin 3 And Human Alpha Defensin 3

As a result of the abuse of antibiotics, the problem of drug-resistant bacteria has become deteriorated. The natural antibacterial peptide has been received many concerns for its strong effect on drug resistant bacteria. Antibacterial peptides are cationic molecules that play an important role in innate immunity and acquired immunity. Defensin is a major member of the antibacterial peptide family, secreted in skin, respiratory tract and other epithelial tissues, and a vital defense against the invasion of exoteric microorganism.In order to construct novel antibacterial fusion gene that has broader or enhanced bioactivity over the original donors, we spliced the genes of bovine neutrophil beta defensin 3 (BNBD3) and human alpha defensin 3 (HNP3) by overlap extension. Meanwhile, the stop codon of BNBD3 and the start codon of HNP3 were deleted. The restriction sites of BamHI and EcoRI were introduced at the two ends of this fusion gene. The reconstructed BNBD3 and HNP3 genes were connected together via a linker sequence encoding Ser-Ser-Gly-Ser-Gly-Ser amino acid residues. The PCR amplified product of the fusion gene was cut by BamHI and EcoRI endonucleases and then cloned into the pGEX-4T-1 expression plasmid...