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A Protocol for Analyzing Hepatitis C Virus Replication
Published on: June 26, 2014
[Molecular cloning and analysis of the 3' terminal sequence of duck hepatitis virus genome]
1Key Laboratory of Preventive Veterinary Medicine of Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100094, China.
Abstract:
The authentic 3' terminal sequences of genomes of duck hepatitis virus (DHV) serotype I strain C80 and serotype I variant strain E63 were obtained by using 3' RACE and RT-PCR techniques. The analysis showed that 3' terminal sequences in the genomes of the two DHV strains include the 3D region of 1359 nucleotides (nt), the terminator codon TGA, and 3'untranslated region (UTR) of 314 nt. The poly (A) tails of C80 and E63 are 18 nt and 19 nt in length respectively. The deduced 3D proteins of 453 amino acids of both DHV strains contain the motifs KDELR, DxxxxD, GxxCSGxxxTxxxNS, YGDD, and FLKR characteristic for RNA polymerase of picornaviruses, which confirms DHV serotype I to be a member of Picornaviridae . The amino acid sequence identities among 3D protein of the two DHV strains with representatives of nine other picornavirus genera range from 16% to 37%, which are within the value ranges (18%-60%) of 3D amino acid sequence identities obtained from inter-genera comparisons. In addition, DHV serotype I possesses the longest 3'UTR in the family Picornaviridae. Phylogenetic analysis of 3D proteins indicated that DHV serotype I may belong to a separated genus of the family Picornaviridae.

