融合海肾荧光素酶的重组病毒在移码和通读研究中的应用初探

<正>海肾荧光素酶(Renilla Luciferase,Rluc)可催化腔肠素(coelenterazine)氧化产生高能量中间产物,氧化过程同时发出蓝色生物荧光,最强荧光波长465 nm。鉴于荧光素酶生物发光的高灵敏度检测特性,由海肾荧光素酶和另一种萤火虫荧光素酶(Firefly Luciferase,Fluc)组成的单报告和双报告基因检测系统广泛应用于生命科学研究领域,如遗传毒性检测、信号通路研究和某些顺式作用元件的

Application of Renilla luciferase-fused virus vector on the study of translational frameshift and readthrough

High sensitive detection system was required for the study of molecular mechanisms involved in translational frameshift or readthrough due to that the translational recoding often occurs with very low efficiency. Although the current dual-luciferase system was sensitive enough to examine the effect of potential elements on frameshift or readthrough, it is not applicable for the study in the context of viral genome. In this study, Renilla luciferase (Rluc)-fused virus vectors were constructed based on the full-length cDNA clones of Tobacco bushy top virus and Beet black scorch virus, respectively. Compared with wild type virus, Rluc-fused TBTV and BBSV increased the value of Rluc by 420 folds and 800 folds respectively, thus providing sufficient sensitivity for the detection of frameshift or readthrough products. Meanwhile, Rluc can function in the context of viral genome. Collectively, engineering the viral genome with the Rluc reporter gene should provide an alternative strategy for exploring the mechanisms of frameshift and readthrough.