摘要
通过构建突变型蛋白表达质粒,将乙型肝炎病毒核心蛋白(Hepatitis B virus core protein, HBc)的赖氨酸位点K96突变为谷氨酰胺(Q)或精氨酸(R),以分别模拟HBc的持续性乙酰化状态以及去乙酰化状态,分析赖氨酸位点K96在病毒衣壳组装过程中的关键性作用;通过建立瞬时转染模型,将野生型及突变型核心蛋白表达质粒转染至细胞中,分别检测突变后的核心蛋白的表达水平和衣壳组装水平,比较分析不同突变对衣壳组装的影响。结果表明:阻断赖氨酸位点K96的乙酰化修饰能够抑制HBc的表达,并且显著降低病毒衣壳的组装水平;而赖氨酸位点K96的持续性乙酰化状态对HBc的表达和病毒衣壳组装水平没有显著影响。
By constructing mutant protein expression plasmid, this study aims to explore the role of lysine residue K96 in viral capsid assembly by mutating this lysine residue into glutamine(Q) or arginine(R), mimic the continuous acetylation or non-acetylation states of HBc, respectively and analyze the key role of lysine residue K96 in the process of viral capsid assembly. The wild-type(WT) and mutant core protein expression plasmids were transfected into cells by establishing a transient transfection model, to detect the expression level of the mutant core proteins and the level of capsid assembly respectively, and compare and analyze the effects of different mutations at this site on the capsid assembly. The results showed that blocking the acetylation of lysine residue K96 can inhibit the expression of HBc and also significantly reduce the level of viral capsid assembly, while the continuous acetylation state of lysine residue K96 had no significant effect on HBc expression or the level of viral capsid assembly.
作者
刘淼雅
刘宽程
梁宗锁
LIU Miaoya;LIU Kuancheng;LIANG Zongsuo(College of Life Sciences and Medicine,Zhejiang Sci-Tech University,Hangzhou 310018,China)
出处
《浙江理工大学学报(自然科学版)》
2022年第1期103-108,共6页
Journal of Zhejiang Sci-Tech University(Natural Sciences)
基金
浙江省自然科学基金项目(LQ19H190003)。