非洲猪瘟病毒p30蛋白的原核表达及其多克隆抗体制备EXPRESSION OF P30 PROTEIN OF AFRICAN SWINE FEVER VIRUS IN PROKARYOTIC CELLS AND PREPARATION OF POLYCLONAL ANTIBODIES
黄剑;徐晶晶;程雪飞;李国新;高飞;童光志;
摘要(Abstract):
本研究利用PCR方法扩增非洲猪瘟病毒(ASFV)SY18株的结构蛋白CP204L基因,并将其克隆至原核表达载体pCold I,构建重组表达质粒pCold I-p30,并将其转化至E.coli BL21(DE3)感受态细胞,经异丙基硫代半乳糖苷(IPTG)诱导表达,SDS-PAGE显示,p30蛋白在大肠杆菌大量表达。获得的p30经纯化免疫小鼠,4次免疫后,采取其血清经过间接免疫荧光和Western blot检测结果表明,获得的多克隆抗体具有明显的特异性。该研究获得的针对p30的多克隆抗体为ASFV早期诊断试及p30蛋白结构功能研究奠定了基础。
关键词(KeyWords): 非洲猪瘟病毒;p30基因;原核表达;多克隆抗体
基金项目(Foundation): 广东省重点领域研发计划项目(2019B020211003);; 国家自然科学基金专项项目(31941017)
作者(Author): 黄剑;徐晶晶;程雪飞;李国新;高飞;童光志;
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