IRES特异性抑制性RNA对HCV IRES介导的蛋白翻译的体外抑制作用

Inhibitory effect of IRES specific inhibitory RNA on HCV IRES mediated protein translation in vitro

目的 研究IRES特异性抑制性RNA(IRNA)对HCVIRES介导蛋白翻译的体外抑制作用。方法 体外化学合成IRNA及其互补体RNA(cIRNA)的cDNA ,以AatⅡ和XbaⅠ双酶切后克隆入具有自剪切作用的顺式核酶载体pGEMRz中 ;最后对重组体的体外转录体在体外无细胞反应体系中的抑制活性进行了初步探讨。结果 IRNA和cIRNA在体外无细胞翻译体系中对HCVIRES介导的蛋白翻译具有相似的抑制活性 ,最大抑制率达 90 %左右。结论 IRNA及cIRNA在体外对HCVIRES介导的蛋白翻译均具有抑制活性。

Aim To study the inhibitory activity of IRES specific inhibitory RNA(IRNA) on HCV IRES mediated protein translation in vitro .Methods In vitro cDNAs of IRNA and complementary IRNA (cIRNA) were chemically synthesized with XbaⅠ overhung at 5'end and AatⅡ at 3'end,and cut with AatⅡ and XbaⅠ together,purified and placed in tripartite ligation reaction with pGEMRz (cut with AatⅡ and XbaⅠ ),which had two different cis -ribozyme site within the multiple enzyme region.Then studied the inhibitory activity of the recombinant transcript in vitro on HCV IRES mediated protein translation in Rabbit Reticulocyte Lysate .Results IRNA and cIRNA had the similar inhibitory activity on HCV IRES mediated protein translation in vitro in Rabbit Reticulocyte Lysate. Compared with the control,the highest inhibitory rate of IRNA was 90 percent.Conclusions IRNA and cIRNA share similar inhibitory activity on HCV IRES mediated protein translation in vitro .