[Expression, purification and antibody preparation of recombinat SARS-CoV X5 protein].
Identifieur interne : 002F65 ( Main/Exploration ); précédent : 002F64; suivant : 002F66[Expression, purification and antibody preparation of recombinat SARS-CoV X5 protein].
Auteurs : Li-Na Wang [République populaire de Chine] ; Jian-Qiang Kong ; Ping Zhu ; Guan-Hua Du ; Wei Wang ; Ke-Di ChengSource :
- Yao xue xue bao = Acta pharmaceutica Sinica [ 0513-4870 ] ; 2008.
Descripteurs français
- KwdFr :
- Animaux, Anticorps (isolement et purification), Corps d'inclusion (), Corps d'inclusion (métabolisme), Données de séquences moléculaires, Escherichia coli (génétique), Escherichia coli (métabolisme), Lapins, Protéines recombinantes (génétique), Protéines recombinantes (immunologie), Protéines recombinantes (isolement et purification), Protéines virales (génétique), Protéines virales (immunologie), Protéines virales (isolement et purification), Régulation de l'expression des gènes viraux, Séquence d'acides aminés, Virus du SRAS (génétique).
- MESH :
- génétique : Escherichia coli, Protéines recombinantes, Protéines virales, Virus du SRAS.
- immunologie : Protéines recombinantes, Protéines virales.
- isolement et purification : Anticorps, Protéines recombinantes, Protéines virales.
- métabolisme : Corps d'inclusion, Escherichia coli.
- Animaux, Corps d'inclusion, Données de séquences moléculaires, Lapins, Régulation de l'expression des gènes viraux, Séquence d'acides aminés.
English descriptors
- KwdEn :
- Amino Acid Sequence, Animals, Antibodies (isolation & purification), Escherichia coli (genetics), Escherichia coli (metabolism), Gene Expression Regulation, Viral, Inclusion Bodies (chemistry), Inclusion Bodies (metabolism), Molecular Sequence Data, Rabbits, Recombinant Proteins (genetics), Recombinant Proteins (immunology), Recombinant Proteins (isolation & purification), SARS Virus (genetics), Viral Proteins (genetics), Viral Proteins (immunology), Viral Proteins (isolation & purification).
- MESH :
- chemical , genetics : Recombinant Proteins, Viral Proteins.
- chemical , immunology : Recombinant Proteins, Viral Proteins.
- chemical , isolation & purification : Antibodies, Recombinant Proteins, Viral Proteins.
- chemistry : Inclusion Bodies.
- genetics : Escherichia coli, SARS Virus.
- metabolism : Escherichia coli, Inclusion Bodies.
- Amino Acid Sequence, Animals, Gene Expression Regulation, Viral, Molecular Sequence Data, Rabbits.
Abstract
X5 protein is one of the putative unknown proteins of SARS-CoV. The recombinant protein has been successfully expressed in E. coli in the form of insoluble inclusion body. The inclusion body was dissolved in high concentration of urea. Affinity Chromatography was preformed to purify the denatured protein, and then the product was refolded in a series of gradient solutions of urea. The purified protein was obtained with the purity of > 95% and the yield of 93.3 mg x L(-1). Polyclonal antibody of this protein was obtained, and Western blotting assay indicated that the X5 protein has the strong property of antigen. Sixty-eight percent of the recombinant protein sequence was confirmed by LC-ESI-MS/MS analysis.
PubMed: 19239038
Affiliations:
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Le document en format XML
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first="Jian-Qiang" last="Kong">Jian-Qiang Kong</name></author><author><name sortKey="Zhu, Ping" sort="Zhu, Ping" uniqKey="Zhu P" first="Ping" last="Zhu">Ping Zhu</name></author><author><name sortKey="Du, Guan Hua" sort="Du, Guan Hua" uniqKey="Du G" first="Guan-Hua" last="Du">Guan-Hua Du</name></author><author><name sortKey="Wang, Wei" sort="Wang, Wei" uniqKey="Wang W" first="Wei" last="Wang">Wei Wang</name></author><author><name sortKey="Cheng, Ke Di" sort="Cheng, Ke Di" uniqKey="Cheng K" first="Ke-Di" last="Cheng">Ke-Di Cheng</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2008">2008</date><idno type="RBID">pubmed:19239038</idno><idno type="pmid">19239038</idno><idno type="wicri:Area/PubMed/Corpus">001954</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001954</idno><idno type="wicri:Area/PubMed/Curation">001954</idno><idno type="wicri:explorRef" wicri:stream="PubMed" 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Laboratory of Biosynthesis of Natural Products, Ministry of Health of PRC, Beijing 100050, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Key Laboratory of Biosynthesis of Natural Products, Ministry of Health of PRC, Beijing 100050</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Kong, Jian Qiang" sort="Kong, Jian Qiang" uniqKey="Kong J" first="Jian-Qiang" last="Kong">Jian-Qiang Kong</name></author><author><name sortKey="Zhu, Ping" sort="Zhu, Ping" uniqKey="Zhu P" first="Ping" last="Zhu">Ping Zhu</name></author><author><name sortKey="Du, Guan Hua" sort="Du, Guan Hua" uniqKey="Du G" first="Guan-Hua" last="Du">Guan-Hua Du</name></author><author><name sortKey="Wang, Wei" sort="Wang, Wei" uniqKey="Wang W" first="Wei" last="Wang">Wei Wang</name></author><author><name sortKey="Cheng, Ke Di" sort="Cheng, Ke Di" uniqKey="Cheng K" first="Ke-Di" last="Cheng">Ke-Di Cheng</name></author></analytic><series><title level="j">Yao xue xue bao = Acta pharmaceutica Sinica</title><idno type="ISSN">0513-4870</idno><imprint><date when="2008" type="published">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence</term><term>Animals</term><term>Antibodies (isolation & purification)</term><term>Escherichia coli (genetics)</term><term>Escherichia coli (metabolism)</term><term>Gene Expression Regulation, Viral</term><term>Inclusion Bodies (chemistry)</term><term>Inclusion Bodies (metabolism)</term><term>Molecular Sequence Data</term><term>Rabbits</term><term>Recombinant Proteins (genetics)</term><term>Recombinant Proteins (immunology)</term><term>Recombinant Proteins (isolation & purification)</term><term>SARS Virus (genetics)</term><term>Viral Proteins (genetics)</term><term>Viral Proteins (immunology)</term><term>Viral Proteins (isolation & purification)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Anticorps (isolement et purification)</term><term>Corps d'inclusion ()</term><term>Corps d'inclusion (métabolisme)</term><term>Données de séquences moléculaires</term><term>Escherichia coli (génétique)</term><term>Escherichia coli (métabolisme)</term><term>Lapins</term><term>Protéines recombinantes (génétique)</term><term>Protéines recombinantes (immunologie)</term><term>Protéines recombinantes (isolement et purification)</term><term>Protéines virales (génétique)</term><term>Protéines virales (immunologie)</term><term>Protéines virales (isolement et purification)</term><term>Régulation de l'expression des gènes viraux</term><term>Séquence d'acides aminés</term><term>Virus du SRAS (génétique)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Recombinant Proteins</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Recombinant Proteins</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Antibodies</term><term>Recombinant Proteins</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Inclusion Bodies</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Escherichia coli</term><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Escherichia coli</term><term>Protéines recombinantes</term><term>Protéines virales</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Protéines recombinantes</term><term>Protéines virales</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Anticorps</term><term>Protéines recombinantes</term><term>Protéines virales</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Escherichia coli</term><term>Inclusion Bodies</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Corps d'inclusion</term><term>Escherichia coli</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Animals</term><term>Gene Expression Regulation, Viral</term><term>Molecular Sequence Data</term><term>Rabbits</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Corps d'inclusion</term><term>Données de séquences moléculaires</term><term>Lapins</term><term>Régulation de l'expression des gènes viraux</term><term>Séquence d'acides aminés</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">X5 protein is one of the putative unknown proteins of SARS-CoV. The recombinant protein has been successfully expressed in E. coli in the form of insoluble inclusion body. The inclusion body was dissolved in high concentration of urea. Affinity Chromatography was preformed to purify the denatured protein, and then the product was refolded in a series of gradient solutions of urea. The purified protein was obtained with the purity of > 95% and the yield of 93.3 mg x L(-1). Polyclonal antibody of this protein was obtained, and Western blotting assay indicated that the X5 protein has the strong property of antigen. Sixty-eight percent of the recombinant protein sequence was confirmed by LC-ESI-MS/MS analysis.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><settlement><li>Pékin</li></settlement></list><tree><noCountry><name sortKey="Cheng, Ke Di" sort="Cheng, Ke Di" uniqKey="Cheng K" first="Ke-Di" last="Cheng">Ke-Di Cheng</name><name sortKey="Du, Guan Hua" sort="Du, Guan Hua" uniqKey="Du G" first="Guan-Hua" last="Du">Guan-Hua Du</name><name sortKey="Kong, Jian Qiang" sort="Kong, Jian Qiang" uniqKey="Kong J" first="Jian-Qiang" last="Kong">Jian-Qiang Kong</name><name sortKey="Wang, Wei" sort="Wang, Wei" uniqKey="Wang W" first="Wei" last="Wang">Wei Wang</name><name sortKey="Zhu, Ping" sort="Zhu, Ping" uniqKey="Zhu P" first="Ping" last="Zhu">Ping Zhu</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Wang, Li Na" sort="Wang, Li Na" uniqKey="Wang L" first="Li-Na" last="Wang">Li-Na Wang</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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