山羊地方性鼻内肿瘤CA蛋白表达及多克隆抗体制备
Journal: Agricultural Science DOI: 10.32629/as.v9i5.3992
Abstract
山羊地方性鼻内肿瘤(enzootic nasal adenocarcinoma, ENA)是由山羊地方性鼻内肿瘤病毒(enzootic nasal tumor virus of goats,ENTV- 2)所引起的一种传染性肿瘤。其在世界范围内广泛存在,对山羊产业造成严重的经济 损失。为获得山羊地方性鼻内肿瘤病毒衣壳蛋白(Capsid protein,CA)及抗CA蛋白的多克隆抗体,本研究根据GenBank中公布的CA基因序列化学 合成CA基因,并连接至pET-30a(+)质粒载体中,构建pET-30a- CA重组原核表达质粒,经测序验证正确后转化大肠杆菌BL21株进行诱导表达,在成功 表达重组蛋白后,使用His- tag亲和层析法纯化蛋白。以纯化的CA重组蛋白作为抗原免疫新西兰大白兔制备多克 隆抗体。使用ELISA方法检测其抗体效价,用Western blot验证多抗的特异性。结果表明,成功在原核表达系统中表达纯化了CA重组蛋白, 原核表达纯化的CA蛋白能够与兔抗CA蛋白多克隆抗体特异性结合。表达的CA蛋白有 较好的抗原性,为制备ENTV-2特异性单克隆抗体及建立血清学方法奠定了基础。
Keywords
山羊地方性鼻内肿瘤；衣壳蛋白；原核表达；多克隆抗体；ELISA检测；Western blot验证
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[16] 刘菲,冯迎春,颜其贵.四川一起山羊地方性鼻内肿瘤疫情的诊断[J].畜牧与兽医, 2011,43(06):83-6.
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[19] ZHAI S-L, LV D-H, XU Z-H, et al. A Novel Enzootic Nasal Tumor Virus Circulating in Goats from Southern China[J].Virus es,2019,11(10):956.
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