The aim of this study is to explore the genomic molecular organization and genogroup of human nomvirus from infected infants in Guangzhou of China. Primers were designed according to the genomic sequence of norovims i...The aim of this study is to explore the genomic molecular organization and genogroup of human nomvirus from infected infants in Guangzhou of China. Primers were designed according to the genomic sequence of norovims in the GenBank, and the nomvirus genome was amplified by RT-PCR. The PCR- products were cloned into T vector and sequenced, and the genomic nucleotide sequences were analyzed with the programs CLUSTAL W/X, DNASTAR and RAT （Recombination Analysis Tool）. The NVgz01 strain genome is 7558 bp in length and encodes three open reading frames （GenBank accession No. is DQ369797）. The genomic sequences of NVgz01 were compared with those of nomvirus in GenBank, which revealed that the homology with genogroup Ⅱ ranges between 76%-90%, and genogroup Ⅰ between 43%-44%. The ORF1 region shared 94% and 88% identity with Mc37 and Famiington strains, respectively; the capsid region （ORF2） shared 65% and 94% identity with Mc37 and Farmington strains, respectively. Phylogenetic trees were reconstructed by the neighbor-joining method. Comparative complete sequence analysis of the NVgz01 with reported human norovirus genomic sequences revealed that this isolate belongs to genogroup Ⅱ . The ORF1 and ORF2 regions shared different identity with Mc37 and Fannington strains, suggesting NVgz01 could be a recombinant virus.展开更多
The capsid protein encoded by ORF2 of human calicivirus(HuCV) expressed in the baculovirus expression system can self-assembled into virus-like particles(VLPs),which would be important for the development of improving...The capsid protein encoded by ORF2 of human calicivirus(HuCV) expressed in the baculovirus expression system can self-assembled into virus-like particles(VLPs),which would be important for the development of improving diagnostic methods.The RT-PCR procedures routinely used to amplify the RNA polymerase region have usually been inefficient for amplification of ORF2,which would bring obstacle for the development of new diagnostic reagents.Through optimization of RT-PCR conditions,we established a simple and high performance RT-PCR method to amplify ORF2,and by which demonstrated a corresponding relationship between the genotype and the antigenic type.It provides conditions for developing diagnostic reagent which accommodates to diagnosis of HuCVs prevailing in China,and also provides the basis for studying the genetic variance and the pathogenesis of HuCV.展开更多
文摘The aim of this study is to explore the genomic molecular organization and genogroup of human nomvirus from infected infants in Guangzhou of China. Primers were designed according to the genomic sequence of norovims in the GenBank, and the nomvirus genome was amplified by RT-PCR. The PCR- products were cloned into T vector and sequenced, and the genomic nucleotide sequences were analyzed with the programs CLUSTAL W/X, DNASTAR and RAT （Recombination Analysis Tool）. The NVgz01 strain genome is 7558 bp in length and encodes three open reading frames （GenBank accession No. is DQ369797）. The genomic sequences of NVgz01 were compared with those of nomvirus in GenBank, which revealed that the homology with genogroup Ⅱ ranges between 76%-90%, and genogroup Ⅰ between 43%-44%. The ORF1 region shared 94% and 88% identity with Mc37 and Famiington strains, respectively; the capsid region （ORF2） shared 65% and 94% identity with Mc37 and Farmington strains, respectively. Phylogenetic trees were reconstructed by the neighbor-joining method. Comparative complete sequence analysis of the NVgz01 with reported human norovirus genomic sequences revealed that this isolate belongs to genogroup Ⅱ . The ORF1 and ORF2 regions shared different identity with Mc37 and Fannington strains, suggesting NVgz01 could be a recombinant virus.
文摘The capsid protein encoded by ORF2 of human calicivirus(HuCV) expressed in the baculovirus expression system can self-assembled into virus-like particles(VLPs),which would be important for the development of improving diagnostic methods.The RT-PCR procedures routinely used to amplify the RNA polymerase region have usually been inefficient for amplification of ORF2,which would bring obstacle for the development of new diagnostic reagents.Through optimization of RT-PCR conditions,we established a simple and high performance RT-PCR method to amplify ORF2,and by which demonstrated a corresponding relationship between the genotype and the antigenic type.It provides conditions for developing diagnostic reagent which accommodates to diagnosis of HuCVs prevailing in China,and also provides the basis for studying the genetic variance and the pathogenesis of HuCV.
