| Summary: | 碩士 === 國立嘉義大學 === 生物科技研究所 === 91 === Internal ribosome entry site (IRES) mediates the initiation of translation of many RNA virus genomes. There were reports indicated that the viral IRES plays important roles in the regulation of viral pathogenesis in the virus hosts. This unusual eukaryotic cap-independent translational mechanism had been employed in development of bi-cistronic expression vector successfully. In this study, we constructed two bi-cistronic expression vectors by the IRES element of enterovirus 71 (EV71) and hepatitis C virus (HCV), named pGS-EV71 and pGS-HCV, respectively.
Both the pGS-EV71 and pGS-HCV containing the β-galactosidase (β-gal) gene and secreted placental alkaline phosphatase (SEAP) gene drive by the human cytomegavirus immediate early promoter (CMV promoter).The expression of the β-gal gene is mediated by cap-dependent translational mechanism and the expression of the SEAP gene is mediated by IRES-mediated translational mechanism. By transient transfection assay in the COS-1 cells, we screened two compounds, A and P20, which could inhibit the EV71-IRES mediated SEAP expression but did not interfere with cap-mediated β-gal expression. However, there were no compounds that can inhibit HCV-IRES mediate translation but not disrupt the cap-dependent translation. These results suggest that the two compounds A and P20 could act as lead compounds to develop the drugs that treat the infection of EV71.
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