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PRRS病毒主要受体蛋白CD151和CD163真核表达载体的构建

DOI: 10.13560/j.cnki.biotech.bull.1985.2014.12.032, PP. 190-194

Keywords: 猪繁殖与呼吸综合征病毒,CD151,分子,CD163,分子,真核表达载体,细胞转染

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Abstract:

为了进一步研究PRRSV与细胞受体之间的相互作用机理,通过基因工程方法从PAM细胞中克隆获得了CD151和CD163全长编码片段,利用SalI和KpnI内切酶对回收得到的CD151(762bp)和CD163(3333bp)DNA片段进行酶切,构建了pIRES2-EGFP-CD151和pIRES2-EGFP-CD163真核表达载体。经PCR和酶切鉴定后,分别提取重组质粒进行细胞转染。荧光显微镜下可见单独转染CD151和CD163基因或CD151/CD163共转染的Marc145细胞表达强烈的绿色荧光,Westernblot分析结果进一步证实,CD151和CD163蛋白在转染后的细胞中获得超表达。猪CD151和CD163真核表达载体的成功构建为进一步探索CD151和CD163在PRRSV感染细胞过程中的协同作用提供了物质基础,特别是对深入研究PRRS病毒的入侵及宿主识别具有重要意义。

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