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cDNA cloning of Korean human norovirus and nucleotidylylation of VPg by norovirus RNA-Dependent RNA polymerase

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Abstract

Norovirus, a member of the Caliciviridae family, is a major causative agent of gastroenteritis worldwide. The cDNA of the entire genome of human norovirus (HuNV) was cloned using the RNA extracted from the stool sample of a Korean patient. The RNA genome consists of 7,559 nucleotides, carries 3 open reading frames (ORFs), 5 and 3 noncoding regions, and a poly(A) tail at the 3 end. Phylogenic analysis of the nucleotide sequence indicated that it belongs to GII.4, the most dominant genogroup. To analyze RNA synthesis and nucleotidylylation of VPg by RNA-dependent RNA polymerase (RdRp), recombinant RdRp and VPg were expressed in Escherichia coli as His-tagged forms. The HuNV RdRp exhibited template and divalent cation-dependent RNA synthesis in vitro. The HuNV RdRp nucleotidylylated HuNV VPg but not murine norovirus (MNV) VPg, whereas MNV RdRp nucleotidylylated both MNV and HuNV VPg more efficiently than HuNV RdRp.

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Authors and Affiliations

  1. Department of Life Science, Sogang University, Seoul, 121-742, Republic of Korea

    Byung Sup Min, Kang Rok Han & Jai Myung Yang

  2. Department of Computer Science & Information Systems, Hanyang Women’s University, Seoul, 133-817, Republic of Korea

    Jung Ihn Lee

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  1. Byung Sup Min
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  2. Kang Rok Han
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  3. Jung Ihn Lee
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  4. Jai Myung Yang
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Correspondence to Jai Myung Yang.

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Min, B.S., Han, K.R., Lee, J.I. et al. cDNA cloning of Korean human norovirus and nucleotidylylation of VPg by norovirus RNA-Dependent RNA polymerase. J Microbiol. 50, 625–630 (2012). https://doi.org/10.1007/s12275-012-2087-4

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  • DOI: https://doi.org/10.1007/s12275-012-2087-4

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