猪流感病毒核衣壳蛋白、非结构蛋白细胞定位的研究.pdfVIP

上海市畜牧兽医学会2011 年学术年会论文集  猪流感病毒核衣壳蛋白、非结构蛋白 细胞定位研究 1, 2 1 1 1, 2 1 李 倩 ，哈 卓 ，师小潇 ，李诗焱 ,刘惠莉 (1.上海市农业科学院畜牧兽医研究所，上海 201106 ；2. 南京农业大学动物医学院，南京 210095) 摘 要：构建真核表达质粒pEGFP-N1-NS1 、pEGFP-N1-NS 、pEGFP-N1-NP 转染MDCK 细胞，以表达的绿色荧光蛋 白(GFP)为标记，观察猪流感病毒NS1 、NS 、NP 蛋白的细胞定位。同时构建真核表达质粒pCAGGS-NS1 、pCAGGS-NS 、 pCAGGS-NP ，转染MDCK 细胞，利用抗NS1 和抗NP 多克隆抗体，进行间接免疫荧光试验，确定重组蛋白的细胞 定位。实验结果表明NS1 、NS 以及NP 蛋白主要定位于细胞核。研究为进一步探讨猪流感病毒NS 、NP 蛋白的生物 学功能奠定基础。 关键词：猪流感病毒；NS 蛋白；NP 蛋白；细胞内定位 CELLULAR LOCALIZATION OF SWINE INFLUENZA VIRUS NS AND NP PROTEIN 1,2 1 1 1 LI Qian , HA Zhuo , SHI Xiao-xiao , LIU Hui-li (Animal Husbandry and Veterinary Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201106,China) Abstract: Eukaryotic expression plasmids pEGFP-N1-NS1, pEGFP-N1-NS, pEGFP-N1-NP were constructed for studying the cellular localization of Swine influenza virus NS and NP proteins. The expressed products of pEGFP-N1-NS1, pEGFP-N1-NS, pEGFP-N1-NP were detected by fluorescence microscope after the recombinant plasmids transfecting into MDCK cells. pCAGGS-NS1, pCAGGS-NS, pCAGGS-NP were constructed and transfect into MDCK cells for protein detection by immunofluorescence analysis using specific polyclonal antibodies. The results show that NS1, NS and NP proteins mainly accumulated in the nucleolus of MDCK cells. The study provides a basis for further research on the proteins’ biological function. Key words: Swine Influenza Virus; NS protein; NP protein; cellular localization 猪流感(swine influenza, SI