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Indexed by:期刊论文
Date of Publication:2016-08-15
Journal:中国农学通报
Included Journals:ISTIC
Volume:32
Issue:23
Page Number:27-31
ISSN No.:1000-6850
Key Words:小反刍兽疫病毒;N蛋白基因;原核表达;间接酶联免疫吸附试验
Abstract:为了获得小反刍兽疫病毒特异性抗原N蛋白,用于血清中抗小反刍兽疫病毒抗体检测方法的建立,根据GenBank中已发表的小反刍兽疫病毒(PPRV)N蛋白核苷酸序列,设计引物,扩增N基因.扩增片段通过Nde Ⅰ和Hind Ⅲ酶切位点插入表达载体pCold Ⅰ.重组蛋白通过pCold Ⅰ载体转化,在大肠杆菌BL21(DE3)中实现了原核表达,经SDS-PAGE和Western Blotting检测表明,表达产物为58 kD的融合蛋白,可被PPRV感染动物血清识别,重组蛋白具有良好的反应原性.用纯化的重组N蛋白作为包被抗原,建立了检测PPRV血清的间接ELISA方法,与国外标准ELISA试剂盒的符合率达98.3％,为建立快速检测小反刍兽疫病毒抗体的检测方法奠定了基础.
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