[Construction of genomic full-length cDNA of SARS coronavirus BJ01 strain and identification of its biological characteristics].
Identifieur interne : 001E96 ( PubMed/Corpus ); 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 ; 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.
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
<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><nlm:affiliation>State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China.</nlm:affiliation></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 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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="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="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" xml:lang="en"><term>Animals</term><term>Chlorocebus aethiops</term><term>Genome, Viral</term><term>Transfection</term><term>Vero Cells</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><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">17302155</PMID><DateCompleted><Year>2009</Year><Month>05</Month><Day>05</Day></DateCompleted><DateRevised><Year>2019</Year><Month>12</Month><Day>10</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0001-6209</ISSN><JournalIssue CitedMedium="Print"><Volume>46</Volume><Issue>6</Issue><PubDate><Year>2006</Year><Month>Dec</Month></PubDate></JournalIssue><Title>Wei sheng wu xue bao = Acta microbiologica Sinica</Title><ISOAbbreviation>Wei Sheng Wu Xue Bao</ISOAbbreviation></Journal><ArticleTitle>[Construction of genomic full-length cDNA of SARS coronavirus BJ01 strain and identification of its biological characteristics].</ArticleTitle><Pagination><MedlinePgn>922-7</MedlinePgn></Pagination><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></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Han</LastName><ForeName>Jian-feng</ForeName><Initials>JF</Initials><AffiliationInfo><Affiliation>State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Jiang</LastName><ForeName>Tao</ForeName><Initials>T</Initials></Author><Author ValidYN="Y"><LastName>Chen</LastName><ForeName>Shui-ping</ForeName><Initials>SP</Initials></Author><Author ValidYN="Y"><LastName>Yu</LastName><ForeName>Man</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Qin</LastName><ForeName>E-de</ForeName><Initials>ED</Initials></Author><Author ValidYN="Y"><LastName>Zhao</LastName><ForeName>Zhuo</ForeName><Initials>Z</Initials></Author><Author ValidYN="Y"><LastName>Li</LastName><ForeName>Xiao-feng</ForeName><Initials>XF</Initials></Author><Author ValidYN="Y"><LastName>Qin</LastName><ForeName>Cheng-feng</ForeName><Initials>CF</Initials></Author><Author ValidYN="Y"><LastName>Deng</LastName><ForeName>Yong-qiang</ForeName><Initials>YQ</Initials></Author><Author ValidYN="Y"><LastName>Zhao</LastName><ForeName>Hui</ForeName><Initials>H</Initials></Author><Author ValidYN="Y"><LastName>Zha</LastName><ForeName>Hai-long</ForeName><Initials>HL</Initials></Author><Author ValidYN="Y"><LastName>Li</LastName><ForeName>Xiao-yu</ForeName><Initials>XY</Initials></Author></AuthorList><Language>chi</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>China</Country><MedlineTA>Wei Sheng Wu Xue Bao</MedlineTA><NlmUniqueID>21610860R</NlmUniqueID><ISSNLinking>0001-6209</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D018076">DNA, Complementary</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002522" MajorTopicYN="N">Chlorocebus aethiops</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018076" MajorTopicYN="N">DNA, Complementary</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D016679" MajorTopicYN="N">Genome, Viral</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D045473" MajorTopicYN="N">SARS Virus</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName><QualifierName UI="Q000472" MajorTopicYN="N">pathogenicity</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014162" MajorTopicYN="N">Transfection</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D014709" MajorTopicYN="N">Vero Cells</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2007</Year><Month>2</Month><Day>17</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2009</Year><Month>5</Month><Day>6</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2007</Year><Month>2</Month><Day>17</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">17302155</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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