title: Development of infectious clones of a wild-type Korean rabies virus and evaluation of their pathogenic potential.
abstract: Most reverse genetic (RG) systems for rabies viruses (RVs) have been constructed on the genome background of laboratory-adapted strains. In this study, we developed an RG system using a Korean wild type (KGH) strain to investigate the pathogenic potential of different strains. We developed a RG system with the KGH strain for the first time. Following the complete genome sequencing of the KGH strain, pKGH infectious clones were constructed using the CMV/T7 promoter, and HamRz and HdvRz were introduced to allow self-cleavage of the synthesized RNA. We successfully recovered the rescued virus by constructing chimeric RVs in which we replaced a part of the construct with the partial gene from the fixed RC-HL strain. The rescued viruses formed clearer and countable plaques in an immunostaining plaque assay, with a distinct plaque morphology. Furthermore, compared with the chimeric RVs, the pKGH/RCinsΔ4 strain containing the KGH strain G protein exhibited a decreased efficiency of cell-to-cell spreading in BHK-21 cells and significantly reduced (100-1000 fold) replication kinetics. However, pKGH/RCinsΔ4 strain-infected mice revealed 100% morbidity at 11days post-infection, whereas other chimeric RV strains showed no mortality. Our RG system is a useful tool for studying differences in the cell-to-cell spreading efficiency and replication with respect to the different internalization patterns of street and fixed laboratory-adapted viruses.
authors.affiliation: Division of Zoonoses, Korea National Research Institute of Health, 187, Osongsaengmyeong 2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, Chungcheongbuk-do, South Korea.
authors.forename: Jun-Sun
authors.initials: JS
authors.lastname: Park
authors.affiliation: Division of Zoonoses, Korea National Research Institute of Health, 187, Osongsaengmyeong 2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, Chungcheongbuk-do, South Korea.
authors.forename: Chi-Kyeong
authors.initials: CK
authors.lastname: Kim
authors.affiliation: Division of Zoonoses, Korea National Research Institute of Health, 187, Osongsaengmyeong 2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, Chungcheongbuk-do, South Korea.
authors.forename: Ji-Hye
authors.initials: JH
authors.lastname: Um
authors.affiliation: Division of Zoonoses, Korea National Research Institute of Health, 187, Osongsaengmyeong 2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, Chungcheongbuk-do, South Korea.
authors.forename: Young Ran
authors.initials: YR
authors.lastname: Ju
authors.affiliation: Division of Zoonoses, Korea National Research Institute of Health, 187, Osongsaengmyeong 2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, Chungcheongbuk-do, South Korea.
authors.forename: Yeong Seon
authors.initials: YS
authors.lastname: Lee
authors.affiliation: Department of Microbiology, College of Medicine and Medical Research Institute,Chungbuk National University, Cheongju-si, South Korea. Electronic address: choiki55@chungbuk.ac.kr.
authors.forename: Young-Ki
authors.initials: YK
authors.lastname: Choi
authors.affiliation: Division of Zoonoses, Korea National Research Institute of Health, 187, Osongsaengmyeong 2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, Chungcheongbuk-do, South Korea. Electronic address: tenksy@korea.kr.
authors.forename: Su Yeon
authors.initials: SY
authors.lastname: Kim
mesh_terms: D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004195:Disease Models, Animal; D019143:Evolution, Molecular; D005260:Female; D023061:Gene Order; D005818:Genetic Engineering; D016679:Genome, Viral; D051379:Mice; D010957:Plasmids / Q000235:genetics; D011818:Rabies / Q000821:virology*; D011820:Rabies virus / Q000235:genetics* / Q000472:pathogenicity; D010948:Viral Plaque Assay; D014774:Virulence; D014779:Virus Replication
chemical_list: D018076:DNA, Complementary
keywords: Fixed virus; KGH strain; RC-HL strain; Rabies virus; Reverse genetic system; Street virus