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本研究在塞内卡病毒A(Senecavirus A,SVA)诱导自噬的基础上，着重探究VP2蛋白在细胞自噬过程中的作用。构建VP2基因真核表达载体pcDNA3.1-VP2，将其转染至PK-15细胞，通过检测自噬蛋白和相关基因的表达情况，明确VP2蛋白对细胞自噬的影响。结果显示，本研究成功构建了pcDNA3.1-VP2真核表达载体，且SVA VP2基因在PK-15细胞中正常表达；与对照组相比，VP2蛋白显著上调LC3蛋白的表达水平（P<0.01）；同时，自噬基因LC3、Beclin-1和ATG5转录水平均显著提高（P<0.01）。综上所述，本研究证实SVA VP2蛋白可诱导PK-15细胞自噬，且VP2蛋白与自噬蛋白和基因表达水平呈正相关，为进一步研究病毒感染与致病机制打下基础。
Abstract：Based on the induction of autophagy by Senecavirus A(SVA),we explored the role of VP2 protein in autophagy. A eukaryotic expression vector of VP2,pcDNA3.1-VP2 was constructed,and transfected into PK-15 cells. The effect of VP2 protein on autophagy was evaluated by measuring expression of autophagy-related proteins and related genes. Results showed that the pcDNA3.1-VP2 eukaryotic expression vector was constructed,and VP2 gene of SVA was expressed in PK-15 cells. Compared with the control group,VP2 protein increased expression of LC3 protein significantly(P<0.01). Simultaneously,transcription of the autophagy-related genes LC3,Beclin-1 and ATG5 was increased significantly(P<0.01). In summary,we demonstrated that the VP2 protein of SVA could induce autophagy in PK-15 cells,and the expression of VP2 protein was positively correlated with expression of autophagy-related proteins and genes. These data lay the foundation for further research on SVA infection and pathogenic mechanisms.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.004068
中图分类号:S852.65
引用信息:
[1]王晶,郭禹,赵云环,等.塞内卡病毒A结构蛋白VP2诱导PK-15细胞自噬的研究[J].病毒学报,2022,38(02):431-438.DOI:10.13242/j.cnki.bingduxuebao.004068.
基金信息:
河北省重点研发计划项目（项目号：19226622D），题目：猪重要新发疫病病原和抗体关键检测技术研究; 河北省农业产业技术体系生猪创新团队（项目号：HBCT2018110207），题目：河北省农业产业技术体系生猪创新团队疫病防控岗~~
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