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猪繁殖与呼吸综合征病毒ORF5/6/7基因真核载体的构建

DOI: 10.7671/j.issn.1001-411X.2012.02.029, PP. 256-258

Keywords: 猪繁殖与呼吸综合征病毒,ORF5/6/7,pEGFP-N1

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Abstract:

针对PRRSVORF5~ORF7设计3对特异性引物,以PRRSVGD-XH株和GD08-1株为模板进行RT-PCR扩增,并将扩增产物克隆到真核表达载体pEGFP-N1中,获得重组质粒pEGFP-N1-(ORF5~ORF7),然后将重组表达质粒转染Marc-145细胞.通过荧光显微镜观察显示,重组质粒pEGFP-N1-(ORF5~ORF7)和载体pEGFP-N1均有绿色荧光出现.结果表明:成功构建了表达PRRSVGD-XH株和PRRSVGD08-1株的GP5、M、N蛋白的真核表达质粒pEGFP-N1-GD-XH-GP、pEGFP-N1-GD-XH-M、pEGFP-N1-GD-XH-N、pEGFP-N1-GD08-1-GP5、pEGFP-N1-GD08-1-M、pEGFP-N1-GD08-1-N.

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