title: Construction and characterization of an infectious cDNA clone of rat hepatitis E virus.
abstract: Rat hepatitis E virus (HEV) is related to human HEV and has been detected in wild rats worldwide. Here, the complete genome of rat HEV strain R63/DEU/2009 was cloned downstream of the T7 RNA polymerase promoter and capped genomic RNA generated by in vitro transcription was injected into nude rats. Rat HEV RNA could be detected in serum and faeces of rats injected intrahepatically, but not in those injected intravenously. Rat HEV RNA-positive faecal suspension was intravenously inoculated into nude rats and Wistar rats leading to rat HEV RNA detection in serum and faeces of nude rats, and to seroconversion in Wistar rats. In addition, rat HEV was isolated in PLC/PRF/5 cells from the rat HEV RNA-positive faecal suspension of nude rats and then passaged. The cell culture supernatant was infectious for nude rats. Genome analysis identified nine point mutations of the cell-culture-passaged virus in comparison with the originally cloned rat HEV genome. The results indicated that infectious rat HEV could be generated from the cDNA clone. As rats are widely used and well-characterized laboratory animals, studies on genetically engineered rat HEV may provide novel insights into organ tropism, replication and excretion kinetics as well as immunological changes induced by hepeviruses.
authors: Department of Virology II, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama, Tokyo 208-0011, Japan.
authors: Tian-Cheng
authors: TC
authors: Li
authors: Department of Clinical Laboratory, Affiliated Hospital of Qingdao University Medical College, Jiangsu Road 16, Qingdao 266003, PR China.
authors: Tingting
authors: T
authors: Yang
authors: Department of Virology II, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama, Tokyo 208-0011, Japan.
authors: Sayaka
authors: S
authors: Yoshizaki
authors: Division of Experimental Animals Research, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama, Tokyo 208-0011, Japan.
authors: Yasushi
authors: Y
authors: Ami
authors: Division of Experimental Animals Research, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama, Tokyo 208-0011, Japan.
authors: Yuriko
authors: Y
authors: Suzaki
authors: Department of Virology II, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama, Tokyo 208-0011, Japan.
authors: Koji
authors: K
authors: Ishii
authors: Department of Virology II, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama, Tokyo 208-0011, Japan.
authors: Kei
authors: K
authors: Haga
authors: Department of Virology II, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama, Tokyo 208-0011, Japan.
authors: Tomofumi
authors: T
authors: Nakamura
authors: Research and Production Technology Department, The Research Foundation for Microbial Diseases of Osaka University, 5-34-4 Kumegawacho Higashimurayama-shi, Tokyo 189-0003, Japan.
authors: Susumu
authors: S
authors: Ochiai
authors: Department of Virology II, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-murayama, Tokyo 208-0011, Japan.
authors: Wakita
authors: W
authors: Takaji
authors: Department Biological Safety, Federal Institute for Risk Assessment, Max-Dohrn-Straße 8-10, 10589 Berlin, Germany.
authors: Reimar
authors: R
authors: Johne
mesh_terms: D000818:Animals; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics*; D005243:Feces / Q000821:virology; D005260:Female; D016752:Hepatitis E virus / Q000235:genetics* / Q000502:physiology*; D007275:Injections, Intravenous; D008297:Male; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics*; D011923:Rats, Nude; D017208:Rats, Wistar; D044967:Serum / Q000821:virology; D014158:Transcription, Genetic; D014773:Virology / Q000379:methods
chemical_list: D018076:DNA, Complementary; D004279:DNA, Viral; D012367:RNA, Viral