文章表达猪传染性胃肠炎病毒TGEV M基因,制备抗M蛋白多克隆抗体。扩增的M基因经Bam H I和Eco R I双酶切后克隆到原核表达载体PGEX-6P-1上,构建重组表达质粒PGEX-6P-1-TGEV-M,经IPTG诱导,获得以包含体形式表达的重组蛋白。将重组蛋白作为免疫原免疫大白兔制备兔抗TGEV-M抗体,进行效价测定及生物活性检测。结果表明,多抗效价达1:262 144;间接ELISA和间接免疫荧光试验说明,此多抗可与TGEV-M重组蛋白及TGEV发生抗原抗体反应。此多抗可作为鉴别诊断试剂将TGEV从众多猪源病毒中区分出来。
【英文摘要】
The aim of this study was to express M gene of porcine transmissible gastroenteritis virus(TGEV) as well as preparation of its specific polyclonal antibodies. The amplified M gene was inserted into the Bam H I-Eco R I site of a prokaryotic expression vector PGEX-6P-1, resulting in a recombinant plasmid PGEX-6P-1-TGEV-M followed by expression in E.coli rossta. The recombinant protein was expressed in the form of inclusion bodies, which was purified and used as immunogen to generate rabbit anti- M polyclonal ...
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