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猪肺炎支原体黏附因子P97蛋白的高效表达与鉴定 被引量:5

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摘要 参考猪肺炎支原体基因组序列设计扩增P97基因片段的一对引物,利用PCR方法扩增出753 bp的基因片段,经测序鉴定正确后,将其定向克隆至原核表达载体pGEX-4T-1中,重组质粒转化大肠杆菌BL21(DE3)表达菌,在IPTG诱导下重组蛋白P97获得表达,通过SDSPAGE及Western blot鉴定重组蛋白的表达情况及蛋白活性。SDS-PAGE结果表明,获得了以包涵体形式表达的重组蛋白,该包涵体蛋白经变性、复性、亲和层析纯化后,Western blot鉴定表明重组蛋白能够被猪肺炎支原体阳性血清特异性识别,具有很好的反应活性与特异性,可以用于P97蛋白单克隆抗体的制备、P97蛋白结构与功能的研究,为进一步开发猪肺炎支原体快速检测试剂盒及新型疫苗研究奠定基础。
出处 《畜牧与兽医》 北大核心 2013年第11期69-72,共4页 Animal Husbandry & Veterinary Medicine
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