Duck hepatitis A virus serotype 1 minigenome: a model for studying the viral 3′UTR effect on viral translation

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Abstract

To date, the genetic replication and translation mechanisms as well as the pathogenesis of duck hepatitis A virus type 1 (DHAV-1) have not been adequately characterized due to the lack of a reliable and efficient cell culture system. Although the full-length infections clone system is the best platform to manipulate the virus, it is relatively difficult to assemble this system due to the lack of a suitable cell line. It has been proven that the minigenome system an efficient reverse genetics system for the study of RNA viruses. In some cases, it can be used to displace the infectious clone of RNA viruses. Here, we generated a minigenome for DHAV-1 with two luciferase reporter genes, firefly luciferase (Fluc) and Renilla luciferase (Rluc). The Rluc gene was used as a reference gene for the normalization of the Fluc gene expression in transfected cells, which provided a platform for studying the regulatory mechanisms of DHAV-1. Furthermore, to investigate the role of DHAV-3′UTR in the regulation of viral protein translation, deletions in the 3′UTR were introduced into the DHAV-1 minigenome. Luciferase activity, an indicator of virus translation, was then determined. These results showed that a minigenome system for DHAV-1 was successfully constructed for the first time and that the complete or partial deletion of the DHAV-3′UTR did not affect the expression level of the reporter gene, indicating that DHAV-1 translation may not be modulated by the viral genomic 3′UTR sequence.

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Liang, R., Li, C., Jin, H., Meng, C., Chen, Z., Zhu, J., … Liu, G. (2015). Duck hepatitis A virus serotype 1 minigenome: a model for studying the viral 3′UTR effect on viral translation. Virus Genes, 51(3), 367–374. https://doi.org/10.1007/s11262-015-1255-0

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