摘要
为研究猪轮状病毒非结构蛋白NSP4的细胞定位及其蛋白功能,将构建的重组质粒pNSP4-EGFP,pviroporin-EGFP和pEGFP-N1空载体质粒转染到SIEC02细胞株,通过与内质网定位蛋白的共聚焦分析、实时荧光定量PCR检测和Western blot分析,发现NSP4主要分布于内质网,少数定位于细胞核;NSP4蛋白可显著上调Bax基因水平(P<0.01)和caspase-3蛋白表达水平。说明猪轮状病毒NSP4蛋白具有类似viroporin的功能,主要定位于内质网上;NSP4依赖或部分依赖caspase的激活和Bax的活化促进细胞凋亡。
The study was to explore the localization of porcine rotavirus nonstructural protein 4（NSP4 ）in the immortalized swine intestinal epithelial cells （SIEC02）established by our laboratory,as well as its pro-tein function.The eukaryotic recombinant plasmids pNSP4-EGFP,pViroporin-EGFP,and pEGFP-N1 vec-tor were transfected transiently into SIEC02 cells.By means of confocal microscopic analysis with endoplas-mic reticulum （ER）positioning vector,real-time fluorescent quantitative PCR and Western blot analysis, the results showed that NSP4 were mainly localized in the ER and few were localized in the cell nucleus;the relative mRNA levels of Bax and the protein expression of caspase-3 were upregulated significantly in the group of NSP4 overexpression （P ＆lt;0.01）.This illustrates that porcine rotavirus NSP4 has similar function with viroporin,mainly localized in the ER;NSP4-induced apoptosis at least partly depended on the caspase and Bax activation in the immortalized swine intestinal epithelial cells.
出处
《动物医学进展》
北大核心
2015年第10期1-4,共4页
Progress In Veterinary Medicine
基金
高等学校博士学科点专项科研基金项目(博导类)(20110204110015)
