title: The pregenome/C RNA of duck hepatitis B virus is not used for translation of core protein during the early phase of infection in vitro.
abstract: Over the course of duck hepatitis B virus (DHBV) replication, one type of RNA (pregenome/C RNA, 3.5 kb) that corresponds to the whole genome of DHBV is generated from the transcription of viral cccDNA. Previous work has proposed three functions for the pregenome/C RNA: it can serve as the pregenome and be packaged into the core protein during the process of replication, and it encodes the mRNA for both the capsid protein and the viral polymerase. However, little is known about the timing of these functions during the different stages of viral infection. In this study, a reverse transcription quantitative real-time PCR assay was developed to analyze the dynamic transcription process of the pregenome/C RNA. The dynamic expression of the core protein was investigated using an indirect immunofluorescence assay (IFA) and by western blot analysis. The generation of pregenome/C RNA began at 12 h post infection and peaked at 20 h post infection; however, the core protein was not detectable until 24h post infection. These results demonstrate that the core protein appeared approximately 12h later than the pregenome/C RNA. These results suggest that the DHBV pregenome/C RNA is not used for the translation of the viral core protein during the early stages of infection.
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, PR China; Avian Disease Research Center, Sichuan Agricultural University, 46 Xinkang Road, Ya'an, Sichuan 625014, PR China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, PR China; Avian Disease Research Center, Sichuan Agricultural University, 46 Xinkang Road, Ya'an, Sichuan 625014, PR China.
authors.forename: Juan
authors.initials: J
authors.lastname: Huang
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, PR China; Avian Disease Research Center, Sichuan Agricultural University, 46 Xinkang Road, Ya'an, Sichuan 625014, PR China; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, Sichuan 611130, PR China). Electronic address: jiary@sicau.edu.cn.
authors.forename: Renyong
authors.initials: R
authors.lastname: Jia
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, PR China; Avian Disease Research Center, Sichuan Agricultural University, 46 Xinkang Road, Ya'an, Sichuan 625014, PR China; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, Sichuan 611130, PR China).
authors.forename: Mingshu
authors.initials: M
authors.lastname: Wang
authors.affiliation: Avian Disease Research Center, Sichuan Agricultural University, 46 Xinkang Road, Ya'an, Sichuan 625014, PR China; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, Sichuan 611130, PR China).
authors.forename: Dekang
authors.initials: D
authors.lastname: Zhu
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, PR China; Avian Disease Research Center, Sichuan Agricultural University, 46 Xinkang Road, Ya'an, Sichuan 625014, PR China; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, Sichuan 611130, PR China).
authors.forename: Shun
authors.initials: S
authors.lastname: Chen
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, PR China; Avian Disease Research Center, Sichuan Agricultural University, 46 Xinkang Road, Ya'an, Sichuan 625014, PR China; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, Sichuan 611130, PR China).
authors.forename: Mafeng
authors.initials: M
authors.lastname: Liu
authors.affiliation: Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, Sichuan 611130, PR China).
authors.forename: Zhongqiong
authors.initials: Z
authors.lastname: Yin
authors.affiliation: Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, Sichuan 611130, PR China).
authors.forename: Yin
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, PR China; Avian Disease Research Center, Sichuan Agricultural University, 46 Xinkang Road, Ya'an, Sichuan 625014, PR China; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu, Sichuan 611130, PR China). Electronic address: chenganchun@vip.163.com.
authors.forename: Anchun
authors.initials: A
authors.lastname: Cheng
mesh_terms: D000818:Animals; D004372:Ducks; D015870:Gene Expression; D016679:Genome, Viral; D018347:Hepadnaviridae Infections; D015611:Hepatitis B Virus, Duck / Q000235:genetics* / Q000378:metabolism; D006524:Hepatitis, Viral, Animal; D014176:Protein Biosynthesis; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism; D013997:Time Factors; D014158:Transcription, Genetic; D014758:Viral Core Proteins / Q000235:genetics* / Q000378:metabolism; D014779:Virus Replication
chemical_list: D012367:RNA, Viral; D014758:Viral Core Proteins
keywords: Core protein; Duck hepatitis B virus; Pregenome/C RNA; RT-qPCR