Diagram of the sub-genomic replicon and reporter plasmid constructs used in the experiments. (A) The HCV replicon is a dicistronic chimeric RNA which contains the gene encoding for neomycin phosphotransferase downstream of the HCV IRES. The second cistern in the same RNA contains EMCV-IRES sequences for efficient translation of HCV non-structural proteins and this chimeric RNA terminates with the HCV 3'UTR. This sub-genomic clone has adaptive mutation S1179I that allows high level replication of RNA in Huh-7 cells and cell colonies develop that are resistant to neomycin. (B) pISRE-Luc reporter plasmid construct containing firefly luciferase reporter gene used to measure IFN-promoter activation in replicon cell lines. This construct has four copies of ISRE sequences positioned up stream of Herpes simplex virus thymidine kinase promoter TATA box, which drives the expression of firefly luciferase. (C) Represents monocistronic reporter constructs that express green fluorescent protein translated by HCV IRES by using a T7 inducible system.
Hazari et al. Virology Journal 2007 4:89 doi:10.1186/1743-422X-4-89