| Title |
cDNA Cloning of Korean Human Norovirus and Nucleotidylylation of VPg by Norovirus RNA-Dependent RNA Polymerase |
| Author |
Byung Sup Min1, Kang Rok Han1, Jung Ihn Lee2, and Jai Myung Yang1* |
| Address |
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 |
| Bibliography |
Journal of Microbiology, 50(4),625-630, 2012,
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| DOI |
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| Key Words |
norovirus, cDNA cloning, RNA-dependent RNA polymerase, VPg |
| 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. |
