title: cDNA cloning of Korean human norovirus and nucleotidylylation of VPg by norovirus RNA-dependent RNA polymerase.
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.
authors.affiliation: Department of Life Science, Sogang University, Seoul 121-742, Republic of Korea.
authors.forename: Byung Sup
authors.initials: BS
authors.lastname: Min
authors.forename: Kang Rok
authors.initials: KR
authors.lastname: Han
authors.forename: Jung Ihn
authors.initials: JI
authors.lastname: Lee
authors.forename: Jai Myung
authors.initials: JM
authors.lastname: Yang
mesh_terms: D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D044466:Asian People; D017250:Caliciviridae Infections / Q000821:virology*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005243:Feces / Q000821:virology; D015870:Gene Expression; D006801:Humans; D029322:Norovirus / Q000235:genetics* / Q000302:isolation & purification; D016366:Open Reading Frames; D012316:RNA Nucleotidyltransferases / Q000378:metabolism; D012333:RNA, Messenger; D012367:RNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D012324:RNA-Dependent RNA Polymerase / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D018076:DNA, Complementary; D004279:DNA, Viral; D012333:RNA, Messenger; D012367:RNA, Viral; D014764:Viral Proteins; D012316:RNA Nucleotidyltransferases; D012324:RNA-Dependent RNA Polymerase