title: Development of a reverse genetics system for a human rabies virus vaccine strain employed in China.
abstract: The CTN rabies virus was isolated from a human in China in 1953 and subsequently attenuated by multiple passaging to a vaccine strain now approved by the WHO. In this study, we describe the development of a reverse genetics system for the CTN rabies virus strain. The recombinant full-length genomic cDNA was flanked by a hammerhead ribozyme (HamRz) and the hepatitis delta virus ribozyme (HdvRz) while the non-coding G-L region was replaced with a green fluorescent protein (GFP) gene. A set of helper plasmids encoding nucleoprotein (N), phosphoprotein (P), and large protein (L) were constructed and co-transfected with recombinant full-length genome plasmid into BHK-21 cells. Recombinant virus was successfully recovered from cloned cDNA under control of the CMV promoter driven by RNA polymerase II. The recombinant virus, CTN-GFP, stably expressed GFP as detected by fluorescence microscopy. A group of 1-day-old suckling mice was challenged with the CTN-GFP strain by intracerebral inoculation, resulting in 100% morbidity and GFP expression was detected in brain tissues. The recombinant virus CTN-GFP strain recovered from cloned cDNA will be useful as a viral vector to express other foreign genes.
authors.affiliation: State Key Laboratory for Molecular Virology and Genetic Engineering, Institute for Viral Disease Control and Prevention, China CDC, 100 Ying Xin St. Xuan Wu District, Beijing 100052, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Huang
authors.forename: Qing
authors.initials: Q
authors.lastname: Tang
authors.forename: Susan A
authors.initials: SA
authors.lastname: Nadin-Davis
authors.forename: Shoufeng
authors.initials: S
authors.lastname: Zhang
authors.forename: Craig D
authors.initials: CD
authors.lastname: Hooper
authors.forename: Pinggang
authors.initials: P
authors.lastname: Ming
authors.forename: Jialiang
authors.initials: J
authors.lastname: Du
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Tao
authors.forename: Rongliang
authors.initials: R
authors.lastname: Hu
authors.forename: Guodong
authors.initials: G
authors.lastname: Liang
mesh_terms: D000818:Animals; D002460:Cell Line; D002681:China; D006224:Cricetinae; D005818:Genetic Engineering / Q000379:methods*; D005827:Genetics, Microbial / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D003698:Hepatitis Delta Virus / Q000201:enzymology / Q000235:genetics; D006801:Humans; D051379:Mice; D010957:Plasmids; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism; D011818:Rabies / Q000821:virology; D011819:Rabies Vaccines / Q000235:genetics* / Q000276:immunology; D011820:Rabies virus / Q000235:genetics* / Q000276:immunology; D016019:Survival Analysis
chemical_list: D016337:RNA, Catalytic; D011819:Rabies Vaccines; D049452:Green Fluorescent Proteins