中国兽医科学2026，Vol.56Issue(9)：1137-1147,11.DOI:10.16656/j.issn.1673-4696.2026.0021
非洲猪瘟病毒跨膜蛋白pX69R的表达及其亚细胞定位分析
Expression and subcellular localization of African swine fever virus transmembrane protein pX69R
摘要
Abstract
The complete X69R gene of African swine fever virus(ASFV)was subcloned into the prokar-yotic expression vector pET-32a(+)by homologous recombination technology.The His-X69R recombinant protein was obtained through IPTG induction and nickel-affinity purification under denaturing condi-tions.The purified His-X69R protein was used to immunize 8-week-old mice to prepare anti-pX69R poly-clonal antibodies.The prepared polyclonal antibodies could specifically recognize FLAG-X69R recom-binant protein expressed in eukaryotic cells and pX69R protein expressed in ASFV-infected porcine alveolar macrophages(PAM).The molecular weight of pX69R protein band in ASFV-infected PAM cells was located at approximately 14 kDa and was primarily expressed in the cytoplasm.Bioinformatics analysis revealed that pX69R lacks a signal peptide,and its amino acids(aa)21-38 constitute a potential trans-membrane domain.Subcellular localization assay indicated that pX69R largely overlapped with endo-plasmic reticulum(ER)signal but showed no significant colocalization with the Golgi apparatus.Trun-cated mutants were further constructed based on the predicted structural domains such as transmembrane region.Subcellular localization analysis demonstrated that deletion of only the 21-38 aa transmem-brane region did not affect ER localization,whereas deletion of 1-41 aa altered its ER localization.These results lay a foundation for further research on the mechanism of pX69R in the replication and pathogenesis of ASFV.关键词
非洲猪瘟病毒/pX69R蛋白/亚细胞定位/内质网定位/多克隆抗体Key words
African swine fever virus/pX69R protein/subcellular localization/ER localization/polyclonal antibody分类
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宣显毅,孙仁杰,张红丽,金子安,徐若楠,田芳琪,赵灵燕,郑肖娟..非洲猪瘟病毒跨膜蛋白pX69R的表达及其亚细胞定位分析[J].中国兽医科学,2026,56(9):1137-1147,11.基金项目
国家重点研发计划项目(2021YFD1801301) （2021YFD1801301）
