Production of egg yolk antibody (IgY) against recombinant porcine epidemic diarrhea virus COE protein into which they were born.
Hu, QingsongeiiaofeiiaofeiLi, LvmuXu, FazhiLv, Yongbiao
Background: Porcine epidemic diarrhea (PED) is a highly contagious disease of pigs, and is characterized by a series of clinical symptoms, such as severe diarrhea, vomiting and dehydration. Partial protective antigen gene (COE gene) of S protein possessing the main B cell epitope, is able to encode proteins with reactogenicity to induce the production of neutralizing antibodies. IgY was found to reduce the mortality in piglets after challenge exposures. Anti-COE IgY antibody has never been reported before, here it is described a method for the production of anti-COE IgY, which could be applied in the treatment for the porcine epidemic diarrhea virus (PEDV) infection.Materials, Methods & Results: A PEDV strain was isolated from a clinical sample. The COE ORF (Open reading frame,ORF) was amplified by PCR and iserted into the pMD18-T clone vector. The isolated was defined as Porcine epidemic diarrhea virus strain JS-HZ2012 subtype by sequencing, the clinical sample was defined as the nucleic acid sequence has a 99.5% homology with that of PEDV CV777 strain. And then the COE ORF was subcloned into pET-32a by T4 DNA ligase and introduced into the E.coli Bal21 (DE3). COE protein was produced by the induction of the E.coli Bal21 containing pET- 32a-COE with isopropyl--D-1-thiogalactopyrannoside (IPTG). Expression of the recombinant COE protein (rCOE) fused with His-tag was analyzTexto completo