title: Recovery of duck hepatitis A virus 3 from a stable full-length infectious cDNA clone.
abstract: Recently, duck hepatitis A virus 3 (DHAV-3) with genetically distinct characteristics from DHAV-1 and DHAV-2 was recognized in South Korea and China. In this short communication, we successfully constructed a stable full-length infectious cDNA clone derived from DHAV-3 by solving instability of cloned full-length cDNA in Escherichia coli (E. coli). The cDNA fragments amplified from the genome of DHAV-3 were assembled and inserted into a low-copy-number plasmid. Finally, a full-length cDNA clone containing an engineered SacII site that served as a genetic marker was obtained. The cDNA clone showed stable by serial passages in E. coli when propagated at 25°C under low level of antibiotic selection. BHK-21 cells were transfected with transcribed RNA from the full-length cDNA clone; infectious viral particles were rescued, showing its fatality to 10-day-old duck embryos. The results indicated that the constructed full-length cDNA clone of DHAV-3 is infectious. By various virological assays, our results indicated that the rescued virus exhibited similar biological properties with the parental virus. Animal experiments revealed that the rescued virus retained the high pathogenicity to 1-day-old ducklings and could induce a fatal hepatitis indistinguishable from its parental virus. Our present studies provide a useful tool for future research on genomic functions and molecular pathogenesis of DHAV-3.
authors.affiliation: Key Laboratory of Animal Epidemiology and Zoonosis of Ministry of Agriculture, College of Veterinary Medicine and State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing, People's Republic of China.
authors.forename: Meng
authors.initials: M
authors.lastname: Pan
authors.forename: Xiaorong
authors.initials: X
authors.lastname: Yang
authors.forename: Jige
authors.initials: J
authors.lastname: Du
authors.forename: Lei
authors.initials: L
authors.lastname: Zhou
authors.forename: Xinna
authors.initials: X
authors.lastname: Ge
authors.forename: Xin
authors.initials: X
authors.lastname: Guo
authors.forename: Jinhua
authors.initials: J
authors.lastname: Liu
authors.forename: Dabing
authors.initials: D
authors.lastname: Zhang
authors.forename: Hanchun
authors.initials: H
authors.lastname: Yang
mesh_terms: D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics*; D004372:Ducks; D004926:Escherichia coli / Q000235:genetics; D005818:Genetic Engineering; D005822:Genetic Vectors; D006524:Hepatitis, Viral, Animal / Q000821:virology; D010849:Picornaviridae / Q000235:genetics* / Q000472:pathogenicity*; D010850:Picornaviridae Infections / Q000662:veterinary / Q000821:virology; D010957:Plasmids; D012367:RNA, Viral / Q000235:genetics*; D016019:Survival Analysis; D014774:Virulence
chemical_list: D018076:DNA, Complementary; D012367:RNA, Viral