鸡Mx-EGFP融合表达蛋白的细胞亚定位及其抗病毒活性研究  

Studies on Subcellular Localization and Anti-viral Activity of the Mx-EGFP Fusion Protein

作　　者：田智泉[1] 倪黎纲 吴晓伟[1] 任丽伟[1] 杨海燕[1] 孙敏[1] 丁爱军[1] 李碧春[1] 

机构地区：[1]扬州大学动物科学与技术学院,扬州225009 [2]江苏省畜牧兽医职业技术学院,泰州225300

出　　处：《畜牧兽医学报》2010年第12期1642-1648,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(30671509;30871791);江苏省高校自然科学重大基础研究项目(08KJA230001);江苏省自然科学基金(BK2007077)资助

摘　　要：为探索鸡Mx基因在细胞中表达的位置,以及Mx-EGFP融合表达时的抗病活性。构建了真核表达载体pcDNA3.0/EGFP-Mx,转染NIH-3T3细胞,通过报告基因EGFP的表达来定位Mx基因在细胞中表达的位置;通过抗VSV病毒试验来研究Mx-EGFP融合蛋白的抗病毒活性。结果显示,融合蛋白大部分在细胞质中表达,而细胞核中也有一部分表达;转染有质粒pcDNA3.0/EGFP-Mx的NIH-3T3细胞感染VSV病毒48h内,细胞没有发生病变;而只转染pcDNA3.0空载体的阴性对照组中,NIH-3T3细胞48h内已经发生病变。研究结果表明,Mx-EGFP融合蛋白主要分布在细胞质中,具有抗VSV病毒活性,能够延缓病毒感染病变时间。This experiment was conducted to study the expression location of chicken Mx gene in cell and the anti-viral activity of the Mx-EGFP fusion protein.The EGFP encoding sequences were subcloned and constructed into the eukaryotic expression vector pcDNA3.0-MMx.Then,NIH-3T3 cells were transfected with the recombinant vector-pcDNA3.0/EGFP-Mx and control vector to observe the subcellular localization of EGFP-Mx.In addition,the fusion protein Mx-EGFP anti-viral activity were studied by anti-VSV virus experiment.The recombinant expression plasmid was ensured after restriction enzyme digestion.Then the recombinant plasmid was transfected into 3T3 cells.Observed under Inverted fluorescence microscope,most of the fusion protein were expressed in the cytoplasm and partly in the nucleus.Attacked with VSV,the cells transfected with only plasmid pcDNA3.0/EGFP-Mx were not pathological changed.But the negative control group transfected with pcDNA3.0 of the NIH-3T3 cells were pathological changed in 48 h.These results indicated that fusion protein was expressed in the cytoplasm mostly and could slow down the time for virus infection lesions.

关 键 词：鸡 MX基因 表达 VSV 

分 类 号：S831.2[农业科学—畜牧学]