[Construction of genomic full-length cDNA of SARS coronavirus BJ01 strain and identification of its biological characteristics].
Identifieur interne : 001E96 ( PubMed/Curation ); précédent : 001E95; suivant : 001E97[Construction of genomic full-length cDNA of SARS coronavirus BJ01 strain and identification of its biological characteristics].
Auteurs : Jian-Feng Han [République populaire de Chine] ; Tao Jiang ; Shui-Ping Chen ; Man Yu ; E-De Qin ; Zhuo Zhao ; Xiao-Feng Li ; Cheng-Feng Qin ; Yong-Qiang Deng ; Hui Zhao ; Hai-Long Zha ; Xiao-Yu LiSource :
- Wei sheng wu xue bao = Acta microbiologica Sinica [ 0001-6209 ] ; 2006.
Descripteurs français
- KwdFr :
- MESH :
- génétique : ADN complémentaire, Virus du SRAS.
- immunologie : Virus du SRAS.
- pathogénicité : Virus du SRAS.
- Animaux, Cellules Vero, Génome viral, Transfection.
English descriptors
- KwdEn :
- MESH :
- chemical , genetics : DNA, Complementary.
- genetics : SARS Virus.
- immunology : SARS Virus.
- pathogenicity : SARS Virus.
- Animals, Chlorocebus aethiops, Genome, Viral, Transfection, Vero Cells.
Abstract
Severe acute respiratory syndrome (SARS) is an important emerging infectious disease which caused by SARS coronavirus (SARS-CoV), and the study of its pathogenesis is needed for the treatment and prevention of this disease. To study the pathogenesis of SARS-CoV using reverse genetics technology, by in vitro ligation using 7 contiguous cDNAs that span the entire genome of the SARS-CoV BJ01 strain, a genomic full-length cDNA was assembled, then using T7 RiboMAX Large Scale RNA Production Systems with the genomic full-length cDNA as template, the RNA transcript was attained. The typical SARS-CoV-resulted cell pathogenic effects were observed when RNA transcript was electroporated into Vero E6 cells. The results of RT-PCR and sequencing of the rescued virus showed that it originated from transcript which derived from the full-length cDNA construct. Rescued virus-infected cells were detected by indirect fluorescent antibody staining demonstrated that it can specifically reaction with SARS-CoV. By CPE method and plaque assay, the titers of the rescued virus and wild-type virus were assessed, which demonstrated there are no significant difference between the viruses and they have similar biological characteristics. Construction of the genomic full-length cDNA of SARS-CoV BJ01 stain successfully and study of the biological characteristics of the rescued virus will provide a useful tool serving for the discovery of molecular pathogenesis and development of candidate vaccines against SARS-CoV.
PubMed: 17302155
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pubmed:17302155Le document en format XML
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<term>SARS Virus (immunology)</term>
<term>SARS Virus (pathogenicity)</term>
<term>Transfection</term>
<term>Vero Cells</term>
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<front><div type="abstract" xml:lang="en">Severe acute respiratory syndrome (SARS) is an important emerging infectious disease which caused by SARS coronavirus (SARS-CoV), and the study of its pathogenesis is needed for the treatment and prevention of this disease. To study the pathogenesis of SARS-CoV using reverse genetics technology, by in vitro ligation using 7 contiguous cDNAs that span the entire genome of the SARS-CoV BJ01 strain, a genomic full-length cDNA was assembled, then using T7 RiboMAX Large Scale RNA Production Systems with the genomic full-length cDNA as template, the RNA transcript was attained. The typical SARS-CoV-resulted cell pathogenic effects were observed when RNA transcript was electroporated into Vero E6 cells. The results of RT-PCR and sequencing of the rescued virus showed that it originated from transcript which derived from the full-length cDNA construct. Rescued virus-infected cells were detected by indirect fluorescent antibody staining demonstrated that it can specifically reaction with SARS-CoV. By CPE method and plaque assay, the titers of the rescued virus and wild-type virus were assessed, which demonstrated there are no significant difference between the viruses and they have similar biological characteristics. Construction of the genomic full-length cDNA of SARS-CoV BJ01 stain successfully and study of the biological characteristics of the rescued virus will provide a useful tool serving for the discovery of molecular pathogenesis and development of candidate vaccines against SARS-CoV.</div>
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<Abstract><AbstractText>Severe acute respiratory syndrome (SARS) is an important emerging infectious disease which caused by SARS coronavirus (SARS-CoV), and the study of its pathogenesis is needed for the treatment and prevention of this disease. To study the pathogenesis of SARS-CoV using reverse genetics technology, by in vitro ligation using 7 contiguous cDNAs that span the entire genome of the SARS-CoV BJ01 strain, a genomic full-length cDNA was assembled, then using T7 RiboMAX Large Scale RNA Production Systems with the genomic full-length cDNA as template, the RNA transcript was attained. The typical SARS-CoV-resulted cell pathogenic effects were observed when RNA transcript was electroporated into Vero E6 cells. The results of RT-PCR and sequencing of the rescued virus showed that it originated from transcript which derived from the full-length cDNA construct. Rescued virus-infected cells were detected by indirect fluorescent antibody staining demonstrated that it can specifically reaction with SARS-CoV. By CPE method and plaque assay, the titers of the rescued virus and wild-type virus were assessed, which demonstrated there are no significant difference between the viruses and they have similar biological characteristics. Construction of the genomic full-length cDNA of SARS-CoV BJ01 stain successfully and study of the biological characteristics of the rescued virus will provide a useful tool serving for the discovery of molecular pathogenesis and development of candidate vaccines against SARS-CoV.</AbstractText>
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