关键词: 猪流行性腹泻病毒;中和抗原表位;转基因玉米;表达

Abstract:

The core neutralizing epitope gene of porcine epidemic diarrhea virus (PEDV\|COE) was expressed in maize（Zea may L.）bioreactor. The COE gene was introduced into the maize inbred line and stable lines highly expressing PEDV\|COE were obtained. Firstly, the coding sequence of the COE gene was optimally modified according to the codon usage bias in maize. Then a synthetic COE gene was cloned into the plant expression vector pBAC9036. Following callus induction, plant regeneration 71 converted regenerated plants were obtained. Through field glyphosate resistance selection, 2 stable T1 transgenic corn lines were obtained.  The result of PCR, RT\|PCR, indirect ELISA and Western blot, etc. indicated that the recombinant COE protein constituted up to 0.122% and 0078% of the total soluble protein was transcripted in the seed of transgenic maize. Finally, the soluble proteins extracted from the seed of  transformed plants were emulsified with Freunds adjuvant and immunized mice by subcutaneous injection route, and the extracts was fed directly to mice for oral immunization. The immunized mice developed specific serum antibody against PEDV comparing with that of the non\|transgenic maize group. These results demonstrated that transgenic maize could not only express the COE polypeptides, but also induce specific antibody against PEDV in mice. Thus, it has laid a solid foundation for R&D of PEDV\|COE transgenic maize vaccine.

Key words: porcine epidemic diarrhea virus, core neutralizing epitope, transgenic maize, expression