title: Duck hepatitis A virus serotype 1 minigenome: a model for studying the viral 3'UTR effect on viral translation.
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.
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, China.|College of Veterinary Medicine, China Agricultural University, Beijing, 100094, China.
authors.forename: Ruiying
authors.initials: R
authors.lastname: Liang
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, China.
authors.forename: Chuanfeng
authors.initials: C
authors.lastname: Li
authors.affiliation: Beijing Institute of Animal Science and Veterinary Medicine, Chinese Academy of Agricultural Sciences, Beijing, 100193, China.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Jin
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, China.
authors.forename: Chunchun
authors.initials: C
authors.lastname: Meng
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, China.
authors.forename: Zongyan
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, China.
authors.forename: Qiuhong
authors.initials: Q
authors.lastname: Miao
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, China.
authors.forename: Chan
authors.initials: C
authors.lastname: Ding
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, China. liugq@shvri.ac.cn.
authors.forename: Guangqing
authors.initials: G
authors.lastname: Liu
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D006224:Cricetinae; D004372:Ducks / Q000821:virology*; D019084:Fluorescent Antibody Technique, Indirect; D017930:Genes, Reporter; D016679:Genome, Viral; D004371:Hepatitis Virus, Duck / Q000235:genetics*; D010957:Plasmids; D014176:Protein Biosynthesis; D012367:RNA, Viral / Q000235:genetics; D017384:Sequence Deletion; D014162:Transfection
chemical_list: D020413:3' Untranslated Regions; D012367:RNA, Viral
keywords: 3′UTR; Duck hepatitis A virus; Minigenome system