Expression Of The Porcine β Defensin 2 In E.coli And Its Antimicrobial Activity Analysis

Defensin is a group of low molecular weight peptides secreted by organism, which is soluble, heat-resistant and so on. It has special antibacterial mechanism against bacteria and showed high antimicrobial activity against a broad range of pathogenic microbiology, including various bacterial, viral, and fungal pathogens , which made it become antimicrobial proteins with broad application . porcineβdefensin 2 is one of 30 porcine defensins with a wide range of antimicrobial activity , Which showed high antibacterial activity. in this research, porcineβ-defensin-2 was cloned and transformed into the prokaryotic expression strain, The target protein was expressed and analyzed. Main contents are as follows:1. Primers were designed according to cDNA sequences published in Genebank , mRNA was extracted from liver of the 35 days pigs. The sequence of pBD2 was obtained by RT-PCR used mRNA as a template. the sequence of matural porcine beta denfensin 2 was obtained using another pair of primers and inserted into expression vector PET30a, Then recombinant vector PET30a-PBD2 was transformed into E. coli BL21 (DE3) PlysS and the expressed strain BL-PET-PBD2 was got.2. The expression strain was induced. The recombinant fusion protein was analyzed by SDS-PAGE and purified by affinity column. Antimicrobial activity analysis showed that recombinant pBD2 fusion protein showed heat-resist and had the activity in suppressing growth of both gram positive bacteria S . aureus and gram negative one E. coli.As described above, the porcine beta defensin 2 was cloned and the recombinant fusion preotein showed antibacterial activity, which is the base for the further experiment...