[Construction of genomic full-length cDNA of SARS coronavirus BJ01 strain and identification of its biological characteristics].
Identifieur interne : 003D81 ( Main/Merge ); précédent : 003D80; suivant : 003D82[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
Links toward previous steps (curation, corpus...)
- to stream PubMed, to step Corpus: 001E96
- to stream PubMed, to step Curation: 001E96
- to stream PubMed, to step Checkpoint: 001F13
- to stream Ncbi, to step Merge: 001872
- to stream Ncbi, to step Curation: 001872
- to stream Ncbi, to step Checkpoint: 001872
Links to Exploration step
pubmed:17302155Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">[Construction of genomic full-length cDNA of SARS coronavirus BJ01 strain and identification of its biological characteristics].</title><author><name sortKey="Han, Jian Feng" sort="Han, Jian Feng" uniqKey="Han J" first="Jian-Feng" last="Han">Jian-Feng Han</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Jiang, Tao" sort="Jiang, Tao" uniqKey="Jiang T" first="Tao" last="Jiang">Tao Jiang</name></author><author><name sortKey="Chen, Shui Ping" sort="Chen, Shui Ping" uniqKey="Chen S" first="Shui-Ping" last="Chen">Shui-Ping Chen</name></author><author><name sortKey="Yu, Man" sort="Yu, Man" uniqKey="Yu M" first="Man" last="Yu">Man Yu</name></author><author><name sortKey="Qin, E De" sort="Qin, E De" uniqKey="Qin E" first="E-De" last="Qin">E-De Qin</name></author><author><name sortKey="Zhao, Zhuo" sort="Zhao, Zhuo" uniqKey="Zhao Z" first="Zhuo" last="Zhao">Zhuo Zhao</name></author><author><name sortKey="Li, Xiao Feng" sort="Li, Xiao Feng" uniqKey="Li X" first="Xiao-Feng" last="Li">Xiao-Feng Li</name></author><author><name sortKey="Qin, Cheng Feng" sort="Qin, Cheng Feng" uniqKey="Qin C" first="Cheng-Feng" last="Qin">Cheng-Feng Qin</name></author><author><name sortKey="Deng, Yong Qiang" sort="Deng, Yong Qiang" uniqKey="Deng Y" first="Yong-Qiang" last="Deng">Yong-Qiang Deng</name></author><author><name sortKey="Zhao, Hui" sort="Zhao, Hui" uniqKey="Zhao H" first="Hui" last="Zhao">Hui Zhao</name></author><author><name sortKey="Zha, Hai Long" sort="Zha, Hai Long" uniqKey="Zha H" first="Hai-Long" last="Zha">Hai-Long Zha</name></author><author><name sortKey="Li, Xiao Yu" sort="Li, Xiao Yu" uniqKey="Li X" first="Xiao-Yu" last="Li">Xiao-Yu Li</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2006">2006</date><idno type="RBID">pubmed:17302155</idno><idno type="pmid">17302155</idno><idno type="wicri:Area/PubMed/Corpus">001E96</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001E96</idno><idno type="wicri:Area/PubMed/Curation">001E96</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">001E96</idno><idno type="wicri:Area/PubMed/Checkpoint">001F13</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001F13</idno><idno type="wicri:Area/Ncbi/Merge">001872</idno><idno type="wicri:Area/Ncbi/Curation">001872</idno><idno type="wicri:Area/Ncbi/Checkpoint">001872</idno><idno type="wicri:doubleKey">0001-6209:2006:Han J:construction:of:genomic</idno><idno type="wicri:Area/Main/Merge">003D81</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">[Construction of genomic full-length cDNA of SARS coronavirus BJ01 strain and identification of its biological characteristics].</title><author><name sortKey="Han, Jian Feng" sort="Han, Jian Feng" uniqKey="Han J" first="Jian-Feng" last="Han">Jian-Feng Han</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Jiang, Tao" sort="Jiang, Tao" uniqKey="Jiang T" first="Tao" last="Jiang">Tao Jiang</name></author><author><name sortKey="Chen, Shui Ping" sort="Chen, Shui Ping" uniqKey="Chen S" first="Shui-Ping" last="Chen">Shui-Ping Chen</name></author><author><name sortKey="Yu, Man" sort="Yu, Man" uniqKey="Yu M" first="Man" last="Yu">Man Yu</name></author><author><name sortKey="Qin, E De" sort="Qin, E De" uniqKey="Qin E" first="E-De" last="Qin">E-De Qin</name></author><author><name sortKey="Zhao, Zhuo" sort="Zhao, Zhuo" uniqKey="Zhao Z" first="Zhuo" last="Zhao">Zhuo Zhao</name></author><author><name sortKey="Li, Xiao Feng" sort="Li, Xiao Feng" uniqKey="Li X" first="Xiao-Feng" last="Li">Xiao-Feng Li</name></author><author><name sortKey="Qin, Cheng Feng" sort="Qin, Cheng Feng" uniqKey="Qin C" first="Cheng-Feng" last="Qin">Cheng-Feng Qin</name></author><author><name sortKey="Deng, Yong Qiang" sort="Deng, Yong Qiang" uniqKey="Deng Y" first="Yong-Qiang" last="Deng">Yong-Qiang Deng</name></author><author><name sortKey="Zhao, Hui" sort="Zhao, Hui" uniqKey="Zhao H" first="Hui" last="Zhao">Hui Zhao</name></author><author><name sortKey="Zha, Hai Long" sort="Zha, Hai Long" uniqKey="Zha H" first="Hai-Long" last="Zha">Hai-Long Zha</name></author><author><name sortKey="Li, Xiao Yu" sort="Li, Xiao Yu" uniqKey="Li X" first="Xiao-Yu" last="Li">Xiao-Yu Li</name></author></analytic><series><title level="j">Wei sheng wu xue bao = Acta microbiologica Sinica</title><idno type="ISSN">0001-6209</idno><imprint><date when="2006" type="published">2006</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Chlorocebus aethiops</term><term>DNA, Complementary (genetics)</term><term>Genome, Viral</term><term>SARS Virus (genetics)</term><term>SARS Virus (immunology)</term><term>SARS Virus (pathogenicity)</term><term>Transfection</term><term>Vero Cells</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN complémentaire (génétique)</term><term>Animaux</term><term>Cellules Vero</term><term>Génome viral</term><term>Transfection</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (immunologie)</term><term>Virus du SRAS (pathogénicité)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA, Complementary</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ADN complémentaire</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="pathogenicity" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="pathogénicité" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Chlorocebus aethiops</term><term>Genome, Viral</term><term>Transfection</term><term>Vero Cells</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Cellules Vero</term><term>Génome viral</term><term>Transfection</term></keywords></textClass></profileDesc></teiHeader><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></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/SrasV1/Data/Main/Merge
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 003D81 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Merge/biblio.hfd -nk 003D81 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= SrasV1
|flux= Main
|étape= Merge
|type= RBID
|clé= pubmed:17302155
|texte= [Construction of genomic full-length cDNA of SARS coronavirus BJ01 strain and identification of its biological characteristics].
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Merge/RBID.i -Sk "pubmed:17302155" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Main/Merge/biblio.hfd \
| NlmPubMed2Wicri -a SrasV1
| This area was generated with Dilib version V0.6.33. |  |