Construction And Identification Of HBD-2 Transgenic Mice And Investigation Of Its Antibacterial Activity In Vivo

Objective The antibacterial activity of HBD-2 was mainly from in vitro antibacterial testing. This study is to construct HBD-2 transgenic mice and evaluate the anti-Staphylococcus aureus capacity of HBD-2 in vivo.Method The full length of HBD-2 cDNA was cloned by RT-PCR from total RNA isolated from SPC-A-1 cells and the pCDNA3.1-HBD2 was constructed. The HBD-2 minigene containing CMV promoter, full length of HBD-2 cDNA, and BGH polyA tail was generated by PCR amplification. The HBD-2 minigenes were introduced into the male pronucli of the fertilized oocytes of C57×ICR hybridized mouse by microinjection and produced offsprings. Integration of HBD-2 gene in the gneomic DNA of offsprings was screened by PCR and HBD-2expression in the tissues of transgenic mice was detected at mRNA levelby RT-PCR and at peptide level by immunohistochemical staining using HBD-2 monoclonal antibody. 1×10~6 clinical isolate of Staphylococcus aureus were intraperitoneally injected and the protective effect of transgenic mice from Staphylocuccus aureus challenge was observed. The bacterial clearing in...