猪流行性腹泻病毒COE核酸疫苗的构建及免疫原性

用疑似患猪流行性腹泻病猪肠病料，扩增部分保护性抗原基因（COE基因），通过T4连接酶将COE基因与真核表达载体plRES2-EGFP进行连接，提取纯化重组质粒。通过脂质体转染vero细胞，用SDS-PAGE和westernblot鉴定COE蛋白的表达。用重组质粒对BALB／c小鼠进行免疫，观察免疫效果。结果显示成功构建了pIRES2-EGFP—COE真核表达载体；目的蛋白在vero细胞中得到表达；重组质粒免疫小鼠能够产生相应抗体。结果表明，COE核酸疫苗可在小鼠体内诱导相应的抗体产生，这为进一步研究猪腹泻病毒的核酸疫苗奠定了基础。

Construction of DNA vaccine of porcine epidemic diarrhea COE and its immuno- genicity in mice

To construct DNA vaccine of COE gene of porcine epidemic diarrhea virus（PEDV） and evaluate its immunogenicity. COE gene of PEDV was amplified and cloned into eukaryotic expres- sion vector pIRES2-EGFP. The recombinant plasmid was purified and transferred into vero cells. SDS-PAGE and Western-blot were used to identify COE protein expressed in vero cells. BALB/c mice were immunized with the recombinant plasmid and immune responses were checked. Results indicated, the eukaryotic expression vector pIRES2-EGFP-COE was successfully constructed,tar- get protein could be expressed in vero cells;and the recombinant plasmid could induce mice to pro- duce responding antibodies. DNA vaccine of COE could induce responding antibodies in mice, which provided a basis for the study of DNA Vaccines against PEDV.