核糖体移码对乙型脑炎病毒基因组表达调控作用解析

The NS1' protein of flavivirus in the Japanese encephalitis virus (JEV) serogroup derives frome -1 ribosomal frameshift in the coding region of NS2A. Currently, the function of NS1' is not completely clear, and the role of ribosomal frameshift in the life cycles of JEV has not been determined. Here, the role of ribosomal frameshift in genome expression of JEV would be studied. The signal sequences for ribosomal frameshift and the coding region of C terminal of NS1' would be changed based on JEV infectious clone pHJEV, respectively. The rescued mutant viruses would be used to infect vero cells and the morphology of plaques formed on vero cells were characterized. Both extracellular and intracellular progeny virus would be analyzed with real time RT-PCR. The amount of IFNs expressed in Ana-1 cells and 3T3-swiss albino cells would be determined during the infection of mutant and parent virus HEN0701. Three-week old mice would be innoculated with mutant virus or HEN0701 to evaluate the virulence of these virus. Complement studies in vitro would be performed to clarify the function of NS1'. These results may clarify the effect of ribosomal frameshift on JEV replication, morphogenesis and virulence.Therefor, the role of ribosomal frameshift in the life cycles of JEV would be elucidated.

乙型脑炎血清群黄病毒通过核糖体-1移码表达NS1′蛋白，但NS1′蛋白功能尚不明确，核糖体移码调控病毒基因组表达对病毒感染复制循环的影响也不清楚。本申请以乙型脑炎病毒为研究对象，操作病毒基因组中核糖体移码信号和改变移码后编码氨基酸序列，测定不同突变病毒核糖体移码效率、空斑形态、感染细胞内外病毒基因组的变化、感染细胞干扰素合成水平及病毒毒力等指标，并以NS1′蛋白体外互补NS1蛋白，探讨核糖体移码对病毒基因组复制、病毒粒子形成及病毒毒力的影响，并研究移码所产生的NS1′蛋白功能，进而明确核糖体移码对JEV基因组表达调控作用及其对病毒感染复制循环的意义。