福建农业学报2025，Vol.40Issue(6)：577-584,8.DOI:10.19303/j.issn.1008-0384.2025.06.005
大黄鱼虹彩病毒ORF 121R蛋白的表达及亚细胞定位
Expression and Subcellular Localization of ORF 121R Protein of Large Yellow Croaker Iridovirus
摘要
Abstract
[Objective]Immunogenicity and subcellular localization of ORF 121R of large yellow croaker iridovirus(LYCIV)were analyzed to understand the pathogenesis role of the protein.[Method]Sequence of LYCIV ORF 121R was compared with those of the homologous proteins,and subcellular localization predicted by Cell-PLoc 2.0.The gene sequence was cloned into pET32a and pEGFP-N1 vectors to construct recombinant plasmids,ORF 121R-His and ORF 121R-EGFP.ORF121R-His was expressed and purified to prepare polyclonal antibodies from mice.Titer and specificity of the antibody were measured using ELISA,western-blotting,and indirect immunofluorescence techniques.After transfecting EPC cells with the ORF 121R-EGFP plasmid,subcellular location of ORF 121R-EGFP was observed under a confocal microscope.[Result]LYCIV ORF 121R was 79.96%homologous in sequence with the immunogenic ISKNV ORF 117R proteins and predicted to localize in the cytoplasm.Successfully constructed from the 1 377 bp orf 121R,the recombinant plasmids were further analyzed.The purified soluble ORF 121R-His had a molecular weight of 72.6 kDa with a polyclonal antibody specifically recognizing natural LYCIV viral protein with a titer of 1:128 000.Whereas ORF 121R-EGFP exhibited a cytoplasmic distribution in EPC cells with the fluorescence signal evenly enriched in the perinuclear region.[Conclusion]LYCIV ORF 121R was mainly found in the cytoplasm.The immunogenicity and specificity of the protein as demonstrated in this study lent it a potential candidate for vaccine production and disease diagnosis on yellow croakers.关键词
大黄鱼虹彩病毒/ORF 121R蛋白/蛋白表达/亚细胞定位Key words
large yellow croaker iridovirus/ORF 121R protein/protein expression/subcellular localization分类
农业科技引用本文复制引用
吴彩霞,刘晓东,池洪树,谢岸桦,陈秀霞,许斌福,郑在予,潘滢,龚晖..大黄鱼虹彩病毒ORF 121R蛋白的表达及亚细胞定位[J].福建农业学报,2025,40(6):577-584,8.基金项目
福建省科技厅对外合作项目(2024I0036) （2024I0036）
福建省科技计划公益类专项(2022R1027004) （2022R1027004）
中央引导地方科技发展专项(2023L3024) （2023L3024）
宁德市揭榜挂帅项目(ND2023A005) （ND2023A005）
