Full-length nucleotide sequence analysis of the S and M segments in Z5 strain of Hantavirus Z5 strain / 中国人兽共患病学报

The M and S segment cDNAs of hantavirus Z5 strain was amplified by RT-PCR,and the purified PCR products were cloned into vector pGEM-T and then sequenced.It was demonstrated that the M genome segment of Z5 was found to be 3 616 nucleotides in length with a single open reading frame encoding 1 135 amino acids.And the S genome segment was 1700 nucleotides in length with a single open reading frame encoding 429 amino acids.As demonstrated by the homologous analysis of nucleotides and amino acids,it was showed that the Z5 strain belonged to hantaan viruses HTN type and was the same subtype of the Z10 strain.It is conclouded that difference in nucleotide sequence exists between Z5 strain with other Hantavirus strains but high level of homology in amino acid sequences is still present.

Molecular cloning and nucleotide sequence analysis of the S-segment in Hantavirus isolated in Zhejiang province / 中国人兽共患病学报

One strain of Hantavirus(HV) was isolated from patients with hemorrhagic fever with renal syndrome (HFRS) in Zhejiang province and its S-segment was cloned and submitted to nucleotide sequence analysis in order to determine the type of strain and extent of genetic variation for further study on its evolution and mutation .The HV antigen was detected from mouse lungs in endemic area by direct immunofluorescene test and the HV-positive sample of mouse lung was inoculated to Vero-E6 cells to isolate virus. The total cellular RNA was extracted from infected cell culture and the S segment gene was amplicated by RT-PCR. Then, the purified PCR product was cloned into T vector for sequencing. The result showed that the full-length sequence of the S segment was 1 700 bp with one open reading frame (ORF) encoding 429 amino acids. Comparison with Hantavirus HTN type showed 85.7%-91.9% homology at the nucleotide level. In comparison with the SEO type of viruses, the homology at nucleotide level was shown to be 71.2%-75%. This new HV strain may be typed as to HTN virus and may be a new subtype of this virus.

Transient Expression of Hantaan Virus S segment Gene Coding region in vitro of Vero-E6 cells / 中国病毒学

The coding region of S genome segment of Hantaan virus (76/118 strain) was inserted into the eukarytic expression plasmidpVR1012. The recombinant expression plasmid pVRS22 was constructed. Vero-E6 cells were transiently transfected in vitro with pVRS22 plasmid. The transient expression of Hantaan virus nucleocapsid proteins in Vero-E6 cells was detected by indirect immunofluorescence assay (IFA) with monoclonal antibody 5H5 against Hantaan virus.