论文部分内容阅读
目的 探讨HCV核心蛋白与HBVX蛋白的协同反式激活作用。方法 构建表达HCV核心蛋白的重组质粒pcDNA3 .1( )core和表达HBVX蛋白的重组质粒pcDNA3 .1( )X ;转染HepG2 细胞 ,从转录和翻译水平鉴定病毒基因的瞬时表达 ;与报告质粒pSV lacZ共转染HepG2 细胞 ,检测 β 半乳糖苷酶表达活性 ,酶的活性反映了表达的肝炎病毒蛋白对SV40病毒早期启动子 增强子功能的影响。结果 构建成HCV核心蛋白及HBVX蛋白的重组表达载体pcDNA3 .1( )core、pcDNA3 .1( )X ;在HepG2 细胞均能瞬时表达相应的肝炎病毒蛋白 ;单独共转染试验中pcDNA3 .1( )core、pcDNA3 .1( )X组的 β 半乳糖苷酶的表达分别是对照的 4.9、3 .5倍 ,两种质粒共同转染时酶的表达是对照的 9倍 ;表达质粒对 β 半乳糖苷酶表达的激活作用呈剂量依赖性。 结论 HepG2 细胞中表达的HCV核心蛋白和HBVX蛋白均具有反式激活SV40早期启动子 增强子的功能 ,并且两种蛋白的反式激活功能具有协同特性。本试验有助于解释HCV、HBV感染 ,尤其是共同感染的致病 癌机制
Objective To investigate the synergistic transactivation of HCV core protein and HBVX protein. Methods Recombinant plasmid pcDNA3.1 () core expressing HCV core protein and pcDNA3.1 () X gene expressing HBVX protein were constructed and transfected into HepG2 cells to detect the transient expression of the virus gene from the transcriptional and translational levels. lacZ was co-transfected into HepG2 cells to detect the expression activity of β-galactosidase, and the activity of the enzyme reflected the effect of the expressed hepatitis virus protein on enhancer function of early promoter of SV40 virus. Results Recombinant expression vector pcDNA3. 1 () core and pcDNA3.1 () X were constructed and transfected into HepG2 cells. The expression of pcDNA3. 1 () ) core and pcDNA3.1 () X group were 4.9, 3.5 times as much as that of the control group, respectively. The expression of the two plasmids was 9 times that of the control The activation of lactosidase expression in a dose-dependent manner. Conclusion Both HCV core protein and HBVX protein expressed in HepG2 cells have the function of transactivating SV40 early promoter enhancer, and the transactivation function of both proteins has synergistic characteristics. This test helps explain the causative mechanisms of HCV and HBV infection, especially co-infections