摘要
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pIRES2-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Western blot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。
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.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2013年第11期1627-1630,共4页
Chinese Journal of Veterinary Science
基金
武汉市财政局创新岗位项目(CX201233)
