SrasV1, Main, Exploration, bibRecord, 002542

Establishment of the eukaryotic cell lines for inducible control of SARS-CoV nucleocapsid gene expression.

Identifieur interne : 002542 ( Main/Exploration ); précédent : 002541; suivant : 002543

Establishment of the eukaryotic cell lines for inducible control of SARS-CoV nucleocapsid gene expression.

Auteurs : Guo-Hui Chang [République populaire de Chine] ; Andrew Dividson ; Lei Lin ; Matt Wilson ; Stuart G. Siddell ; Qing-Yu Zhu

Source :

Virologica Sinica [ 1995-820X ] ; 2010.

RBID : pubmed:20960182

Descripteurs français

KwdFr :
- Analyse de séquence, Animaux, Biologie informatique, Cricetinae, Doxycycline (métabolisme), Expression des gènes, Lignée cellulaire, Plasmides, Protéines nucléocapside (biosynthèse), Protéines nucléocapside (génétique), Régulation de l'expression des gènes, Vecteurs génétiques, Virus du SRAS (génétique).
MESH :
- biosynthèse : Protéines nucléocapside.
- génétique : Protéines nucléocapside, Virus du SRAS.
- métabolisme : Doxycycline.
- Analyse de séquence, Animaux, Biologie informatique, Cricetinae, Expression des gènes, Lignée cellulaire, Plasmides, Régulation de l'expression des gènes, Vecteurs génétiques.

English descriptors

KwdEn :
- Animals, Cell Line, Computational Biology, Cricetinae, Doxycycline (metabolism), Gene Expression, Gene Expression Regulation, Genetic Vectors, Nucleocapsid Proteins (biosynthesis), Nucleocapsid Proteins (genetics), Plasmids, SARS Virus (genetics), Sequence Analysis.
MESH :
- chemical , biosynthesis : Nucleocapsid Proteins.
- chemical , genetics : Nucleocapsid Proteins.
- chemical , metabolism : Doxycycline.
- genetics : SARS Virus.
- Animals, Cell Line, Computational Biology, Cricetinae, Gene Expression, Gene Expression Regulation, Genetic Vectors, Plasmids, Sequence Analysis.

Abstract

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.

DOI: 10.1007/s12250-010-3124-2
PubMed: 20960182

Affiliations:

République populaire de Chine

Le document en format XML

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<front><div type="abstract" xml:lang="en">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.</div>
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Establishment of the eukaryotic cell lines for inducible control of SARS-CoV nucleocapsid gene expression.

Establishment of the eukaryotic cell lines for inducible control of SARS-CoV nucleocapsid gene expression.

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

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Pour générer des pages wiki

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