cDNA Cloning of Korean Human Norovirus and Nucleotidylylation of VPg by Norovirus RNA-Dependent RNA Polymerase

Byung Sup Min; Kang Rok Han; Jung Ihn Lee; Jai Myung Yang

doi:10.1007/s12275-012-2087-4

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cDNA Cloning of Korean Human Norovirus and Nucleotidylylation of VPg by Norovirus RNA-Dependent RNA Polymerase: Byung Sup Min ¹, Kang Rok Han ¹, Jung Ihn Lee ², Jai Myung Yang ¹; Journal of Microbiology 2012;50(4):625-630.
DOI: https://doi.org/10.1007/s12275-012-2087-4
Published online: August 25, 2012

¹Department of Life Science, Sogang University, Seoul 121-742, Republic of Korea, ²Department of Computer Science & Information Systems, Hanyang Women’s University, Seoul 133-817, Republic of Korea

Corresponding author: Jai Myung Yang , Tel: +82-2-705-8457,

Received: 16 February 2012 • Accepted: 29 May 2012

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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.

Keywords: norovirus;cDNA cloning;RNA-dependent RNA polymerase;VPg

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cDNA Cloning of Korean Human Norovirus and Nucleotidylylation of VPg by Norovirus RNA-Dependent RNA Polymerase

J. Microbiol. 2012;50(4):625-630. Published online August 25, 2012

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

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