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猪繁殖与呼吸综合征病毒各结构蛋白的融合表达
Fusion expression of structural proteins of porcine reproductive and respiratory syndrome virus
【摘要】 本研究以测序质粒为模板,扩增出结构蛋白基因ORF2a-ORF7,将此基因片段插入真核表达载体pEGFP-N3中构建重组质粒pEGFP-N3-(ORF2a～ORF7),然后将重组表达质粒转染293T细胞。通过荧光显微镜观察显示,重组质粒pEGFP-N3-(ORF2a～ORF7)和载体pEGFP-N3均有绿色荧光出现。SDS-PAGE电泳和western blot分析结果表明重组质粒pEGFP-N3-(ORF2a～ORF7)和载体pEGFP-N3均有EGFP表达;仅ORF7有43ku的融合蛋白EGFP出现,与预期大小相符;其他重组质粒pEGFP-N3-(ORF2a～ORF6)均没有融合蛋白出现。本实验为进一步研究这些蛋白的结构和生物学功能奠定了基础。
【Abstract】 In this report, the structural protein genes of porcine reproductive and respiratory syndrome virus(PRRSV)(ORF2a-ORF7) were PCR amplified and cloned into eukaryotic expressing vector pEGFP-N3.The recombinant plasmid pEGFP-N3-(ORF2a-ORF7) was transfected into 293T cell.Fluorescence microscopy showed that EGFP were expressed in all transfected cells.SDS-PAGE and western-blot indicated that only the ORF7 gene was successfully expressed.
- 【文献出处】 中国预防兽医学报 ,Chinese Journal of Preventive Veterinary Medicine , 编辑部邮箱 ,2009年02期
- 【分类号】S852.659.6
- 【被引频次】6
- 【下载频次】196