Source:http://linkedlifedata.com/resource/pubmed/id/20960182
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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
5
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pubmed:dateCreated |
2010-10-20
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pubmed:abstractText |
In order to establish the eukaryotic cell lines for inducible control of SARS-CoV nucleocapsid gene expression. The recombinant plasmid of pTRE-Tight-SARS-N was constructed by using the plasmid p8S as the PCR template which contains a cDNA clone covering the nucleocapsid gene of SARS-CoV HKU-39449. Restriction enzymes digestion and sequence analysis indicated the recombinant plasmid of pTRE-Tight-SARS-N contained the nucleocapsid gene with the optimized nucleotide sequence which will improve the translation efficiency. Positive cell clones were selected by cotransfecting pTRE-Tight-SARS-N with the linear marker pPUR to BHK-21 Tet-on cells in the presence of puromycin. A set of double-stable eukaryotic cell lines (BHK-Tet-SARS-N) with inducible control of the SARS-CoV neucleocapsid gene expression was identified by using SDS-PAGE and Western-blot analysis. The expression of SARS-CoV nucleocapsid protein was tightly regulated by the varying concentration of doxcycline in the constructed double-stable cell line. The constructed BHK-Tet-SARS-N cell strains will facilitate the rescue of SARS-CoV in vitro and the further reverse genetic research of SARS-CoV.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Oct
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pubmed:issn |
1995-820X
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pubmed:author | |
pubmed:issnType |
Electronic
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pubmed:volume |
25
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
361-8
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pubmed:meshHeading |
pubmed-meshheading:20960182-Animals,
pubmed-meshheading:20960182-Cell Line,
pubmed-meshheading:20960182-Computational Biology,
pubmed-meshheading:20960182-Cricetinae,
pubmed-meshheading:20960182-Doxycycline,
pubmed-meshheading:20960182-Gene Expression,
pubmed-meshheading:20960182-Gene Expression Regulation,
pubmed-meshheading:20960182-Genetic Vectors,
pubmed-meshheading:20960182-Nucleocapsid Proteins,
pubmed-meshheading:20960182-Plasmids,
pubmed-meshheading:20960182-SARS Virus,
pubmed-meshheading:20960182-Sequence Analysis
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pubmed:year |
2010
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pubmed:articleTitle |
Establishment of the eukaryotic cell lines for inducible control of SARS-CoV nucleocapsid gene expression.
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pubmed:affiliation |
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100071, China. changguohui999@yahoo.com.cn
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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