【摘要】 为了研究猪干扰素刺激基因20(interferon-stimulated gene 20,ISG20)的序列信息及其表达情况，并探讨其对猪流行性腹泻病毒(Porcine epidemic diarrhea virus, PEDV)增殖的影响，以提取的猪小肠上皮细胞IPEC-J2总RNA为模板扩增猪ISG20基因的CDS区序列，并应用生物信息学进行分析，进而构建pcDNA3.1-3×Flag-ISG20重组质粒转染入IPEC-J2细胞鉴定表达情况。此外，在转染重组质粒的细胞中感染PEDV分析ISG20对PEDV增殖的影响。结果表明，猪ISG20基因CDS区的大小为516 bp,编码171个氨基酸，理论等电点(pI)为8.84;猪ISG20存在2个N-糖基化位点和23个磷酸化位点，主要定位于细胞骨架和细胞质中，以α-螺旋和无规则卷曲构成；构建的pcDNA3.1-3×Flag-ISG20重组质粒能够在IPEC-J2细胞中表达，蛋白大小约为19.6 ku,而且可以明显抑制PEDV的增殖；克隆了猪ISG20基因，并且在IPEC-J2细胞中重组质粒pcDNA3.1-3×Flag-ISG20能够表达且发挥抑制PEDV增殖的作用，研究结果为深入探讨PEDV与ISG20的相互作用，以及以此为靶点筛选抗PEDV生物制剂提供了思路。更多还原

【Abstract】 This article aims to study the sequence information and expression of the porcine interferon stimulated gene 20(ISG20),and to explore its impact on the proliferation of porcine epizootic diarrhea virus(PEDV),using the extracted total RNA of porcine small intestine epithelial cells IPEC-J2 as a template to amplify the CDS region sequence of porcine ISG20 gene, and applying bioinformatics analysis to construct pcDNA3.1-3×Flag-ISG20 recombinant plasmid, the expression was identified by transfection into IPEC-J2 cells.In addition, the effect of ISG20 on PEDV proliferation in cells was analyzed after transfection with recombinant plasmids.The results showed that the size of the CDS region of the porcine ISG20 gene was 516bp, encoding 171 amino acids, and the theoretical isoelectric point(pI) was 8.84; Porcine ISG20 has 2 N-glycosylation sites and 23 phosphorylation sites, mainly located in the cytoskeleton and cytoplasm, which composed of α-spirals and irregular curls.The pcDNA3.1-3×Flag-ISG20 recombinant plasmid can be expressed in IPEC-J2 cells, with a protein size of approximately 19.6 ku, and can significantly inhibit the proliferation of PEDV.In summary, the results of this study cloned the porcine ISG20 gene, and recombinant plasmid pcDNA3.1-3 in IPEC-J2 cells×Flag-ISG20 can be expressed and exert an inhibitory effect on PEDV proliferation.This study provides insights into the interaction between PEDV and ISG20,as well as the screening of anti PEDV biologics using this as a target.更多还原