The expression of SARS–CoV M gene in P. Pastoris and the diagnostic utility of the expression product
Identifieur interne : 005020 ( Main/Exploration ); précédent : 005019; suivant : 005021The expression of SARS–CoV M gene in P. Pastoris and the diagnostic utility of the expression product
Auteurs : Xueqing Han [République populaire de Chine] ; Mark Bartlam [République populaire de Chine] ; Ying-Hua Jin [République populaire de Chine] ; Xiangtao Liu [République populaire de Chine] ; Xiaojing He [République populaire de Chine] ; Xuepeng Cai [République populaire de Chine] ; Qingqe Xie [République populaire de Chine] ; Zihe Rao [République populaire de Chine]Source :
- Journal of Virological Methods [ 0166-0934 ] ; 2004.
Descripteurs français
- KwdFr :
- Adulte, Alignement de séquences, Anticorps antiviraux (sang), Clonage moléculaire, Données de séquences moléculaires, Femelle, Gènes viraux, Humains, Mâle, Pichia (génétique), Pichia (métabolisme), Protéines de la matrice virale (biosynthèse), Protéines de la matrice virale (génétique), Protéines de la matrice virale (immunologie), Protéines de la matrice virale (isolement et purification), Protéines recombinantes (biosynthèse), Syndrome respiratoire aigu sévère (diagnostic), Syndrome respiratoire aigu sévère (virologie), Séquence d'acides aminés, Technique de Western, Techniques immunologiques, Test ELISA, Vecteurs génétiques, Électrophorèse sur gel de polyacrylamide.
- MESH :
- biosynthèse : Protéines de la matrice virale, Protéines recombinantes.
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : Pichia, Protéines de la matrice virale.
- immunologie : Protéines de la matrice virale.
- isolement et purification : Protéines de la matrice virale.
- métabolisme : Pichia.
- sang : Anticorps antiviraux.
- virologie : Syndrome respiratoire aigu sévère.
- Adulte, Alignement de séquences, Clonage moléculaire, Données de séquences moléculaires, Femelle, Gènes viraux, Humains, Mâle, Séquence d'acides aminés, Technique de Western, Techniques immunologiques, Test ELISA, Vecteurs génétiques, Électrophorèse sur gel de polyacrylamide.
English descriptors
- KwdEn :
- Adult, Amino Acid Sequence, Antibodies, Viral (blood), Blotting, Western, Cloning, Molecular, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay, Female, Genes, Viral, Genetic Vectors, Humans, Immunologic Techniques, Male, Molecular Sequence Data, Pichia (genetics), Pichia (metabolism), Recombinant Proteins (biosynthesis), Sequence Alignment, Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (virology), Viral Matrix Proteins (biosynthesis), Viral Matrix Proteins (genetics), Viral Matrix Proteins (immunology), Viral Matrix Proteins (isolation & purification).
- MESH :
- chemical , biosynthesis : Recombinant Proteins, Viral Matrix Proteins.
- chemical , blood : Antibodies, Viral.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : Pichia, Viral Matrix Proteins.
- chemical , immunology : Viral Matrix Proteins.
- chemical , isolation & purification : Viral Matrix Proteins.
- metabolism : Pichia.
- virology : Severe Acute Respiratory Syndrome.
- Adult, Amino Acid Sequence, Blotting, Western, Cloning, Molecular, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay, Female, Genes, Viral, Genetic Vectors, Humans, Immunologic Techniques, Male, Molecular Sequence Data, Sequence Alignment.
Abstract
High-level protein expression is an important means of obtaining large amounts of viral proteins to investigate further their biological properties. To express the membrane (M) protein of SARS–CoV at high-level in vitro, the
Url:
DOI: 10.1016/j.jviromet.2004.08.015
PubMed: 15488627
PubMed Central: 7112794
Affiliations:
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Le document en format XML
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Pastoris</italic> and the diagnostic utility of the expression product</title><author><name sortKey="Han, Xueqing" sort="Han, Xueqing" uniqKey="Han X" first="Xueqing" last="Han">Xueqing Han</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Structural Biology, Tsinghua University and National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Science, Beijing, PR China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Laboratory of Structural Biology, Tsinghua University and National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Science, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation><affiliation wicri:level="3"><nlm:aff id="aff2">Lanzhou Institute of Veterinary Medicine, Chinese Academy of Agricultural Sciences, Baishiquio Road 30, Beijing, PR China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Lanzhou Institute of Veterinary Medicine, Chinese Academy of Agricultural Sciences, Baishiquio Road 30, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Bartlam, Mark" sort="Bartlam, Mark" uniqKey="Bartlam M" first="Mark" last="Bartlam">Mark Bartlam</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Structural Biology, Tsinghua University and National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Science, Beijing, PR China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Laboratory of Structural Biology, Tsinghua University and National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Science, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Jin, Ying Hua" sort="Jin, Ying Hua" uniqKey="Jin Y" first="Ying-Hua" last="Jin">Ying-Hua Jin</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Structural Biology, Tsinghua University and National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Science, Beijing, PR China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Laboratory of Structural Biology, Tsinghua University and National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Science, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Liu, Xiangtao" sort="Liu, Xiangtao" uniqKey="Liu X" first="Xiangtao" last="Liu">Xiangtao Liu</name><affiliation wicri:level="3"><nlm:aff id="aff2">Lanzhou Institute of Veterinary Medicine, Chinese Academy of Agricultural Sciences, Baishiquio Road 30, Beijing, PR China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Lanzhou Institute of Veterinary Medicine, Chinese Academy of Agricultural Sciences, Baishiquio Road 30, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="He, Xiaojing" sort="He, Xiaojing" uniqKey="He X" first="Xiaojing" last="He">Xiaojing He</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Structural Biology, Tsinghua University and National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Science, Beijing, PR China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Laboratory of Structural Biology, Tsinghua University and National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Science, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Cai, Xuepeng" sort="Cai, Xuepeng" uniqKey="Cai X" first="Xuepeng" last="Cai">Xuepeng Cai</name><affiliation wicri:level="3"><nlm:aff id="aff2">Lanzhou Institute of Veterinary Medicine, Chinese Academy of Agricultural Sciences, Baishiquio Road 30, Beijing, PR China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Lanzhou Institute of Veterinary Medicine, Chinese Academy of Agricultural Sciences, Baishiquio Road 30, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Xie, Qingqe" sort="Xie, Qingqe" uniqKey="Xie Q" first="Qingqe" last="Xie">Qingqe Xie</name><affiliation wicri:level="3"><nlm:aff id="aff2">Lanzhou Institute of Veterinary Medicine, Chinese Academy of Agricultural Sciences, Baishiquio Road 30, Beijing, PR China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Lanzhou Institute of Veterinary Medicine, Chinese Academy of Agricultural Sciences, Baishiquio Road 30, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Rao, Zihe" sort="Rao, Zihe" uniqKey="Rao Z" first="Zihe" last="Rao">Zihe Rao</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Structural Biology, Tsinghua University and National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Science, Beijing, PR China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Laboratory of Structural Biology, Tsinghua University and National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Science, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author></analytic><series><title level="j">Journal of Virological Methods</title><idno type="ISSN">0166-0934</idno><idno type="eISSN">1879-0984</idno><imprint><date when="2004">2004</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Adult</term><term>Amino Acid Sequence</term><term>Antibodies, Viral (blood)</term><term>Blotting, Western</term><term>Cloning, Molecular</term><term>Electrophoresis, Polyacrylamide Gel</term><term>Enzyme-Linked Immunosorbent Assay</term><term>Female</term><term>Genes, Viral</term><term>Genetic Vectors</term><term>Humans</term><term>Immunologic Techniques</term><term>Male</term><term>Molecular Sequence Data</term><term>Pichia (genetics)</term><term>Pichia (metabolism)</term><term>Recombinant Proteins (biosynthesis)</term><term>Sequence Alignment</term><term>Severe Acute Respiratory Syndrome (diagnosis)</term><term>Severe Acute Respiratory Syndrome (virology)</term><term>Viral Matrix Proteins (biosynthesis)</term><term>Viral Matrix Proteins (genetics)</term><term>Viral Matrix Proteins (immunology)</term><term>Viral Matrix Proteins (isolation & purification)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Adulte</term><term>Alignement de séquences</term><term>Anticorps antiviraux (sang)</term><term>Clonage moléculaire</term><term>Données de séquences moléculaires</term><term>Femelle</term><term>Gènes viraux</term><term>Humains</term><term>Mâle</term><term>Pichia (génétique)</term><term>Pichia (métabolisme)</term><term>Protéines de la matrice virale (biosynthèse)</term><term>Protéines de la matrice virale (génétique)</term><term>Protéines de la matrice virale (immunologie)</term><term>Protéines de la matrice virale (isolement et purification)</term><term>Protéines recombinantes (biosynthèse)</term><term>Syndrome respiratoire aigu sévère (diagnostic)</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Séquence d'acides aminés</term><term>Technique de Western</term><term>Techniques immunologiques</term><term>Test ELISA</term><term>Vecteurs génétiques</term><term>Électrophorèse sur gel de polyacrylamide</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Recombinant Proteins</term><term>Viral Matrix Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>Antibodies, Viral</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Protéines de la matrice virale</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Pichia</term><term>Viral Matrix Proteins</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Pichia</term><term>Protéines de la matrice virale</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Protéines de la matrice virale</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Viral Matrix Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Viral Matrix Proteins</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Protéines de la matrice virale</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Pichia</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Pichia</term></keywords><keywords scheme="MESH" qualifier="sang" xml:lang="fr"><term>Anticorps antiviraux</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Adult</term><term>Amino Acid Sequence</term><term>Blotting, Western</term><term>Cloning, Molecular</term><term>Electrophoresis, Polyacrylamide Gel</term><term>Enzyme-Linked Immunosorbent Assay</term><term>Female</term><term>Genes, Viral</term><term>Genetic Vectors</term><term>Humans</term><term>Immunologic Techniques</term><term>Male</term><term>Molecular Sequence Data</term><term>Sequence Alignment</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Adulte</term><term>Alignement de séquences</term><term>Clonage moléculaire</term><term>Données de séquences moléculaires</term><term>Femelle</term><term>Gènes viraux</term><term>Humains</term><term>Mâle</term><term>Séquence d'acides aminés</term><term>Technique de Western</term><term>Techniques immunologiques</term><term>Test ELISA</term><term>Vecteurs génétiques</term><term>Électrophorèse sur gel de polyacrylamide</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>High-level protein expression is an important means of obtaining large amounts of viral proteins to investigate further their biological properties. To express the membrane (M) protein of SARS–CoV at high-level in vitro, the <italic>M</italic> gene fragment was amplified and cloned it into the <italic>Pichia Pastoris</italic> expression vector pPICZαA. SDS–PAGE and Western blotting analysis of the induced products of recombinant yeast transformant indicated that successful high-level expression of M protein was achieved, and that the expression product was similar antigenically to the natural protein. Purified recombinant M protein was used subsequently as an ELISA antigen for detection of eight serum samples screened previously by whole virus ELISA and immunofluorescence assay, and consistent results were obtained. These findings suggest that the recombinant M protein may be useful as a diagnostic reagent.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Corse, E" uniqKey="Corse E">E. Corse</name></author><author><name sortKey="Machamaer, C E" uniqKey="Machamaer C">C.E. 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Cao</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>République populaire de Chine</li></country><settlement><li>Pékin</li></settlement></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Han, Xueqing" sort="Han, Xueqing" uniqKey="Han X" first="Xueqing" last="Han">Xueqing Han</name></noRegion><name sortKey="Bartlam, Mark" sort="Bartlam, Mark" uniqKey="Bartlam M" first="Mark" last="Bartlam">Mark Bartlam</name><name sortKey="Cai, Xuepeng" sort="Cai, Xuepeng" uniqKey="Cai X" first="Xuepeng" last="Cai">Xuepeng Cai</name><name sortKey="Han, Xueqing" sort="Han, Xueqing" uniqKey="Han X" first="Xueqing" last="Han">Xueqing Han</name><name sortKey="He, Xiaojing" sort="He, Xiaojing" uniqKey="He X" first="Xiaojing" last="He">Xiaojing He</name><name sortKey="Jin, Ying Hua" sort="Jin, Ying Hua" uniqKey="Jin Y" first="Ying-Hua" last="Jin">Ying-Hua Jin</name><name sortKey="Liu, Xiangtao" sort="Liu, Xiangtao" uniqKey="Liu X" first="Xiangtao" last="Liu">Xiangtao Liu</name><name sortKey="Rao, Zihe" sort="Rao, Zihe" uniqKey="Rao Z" first="Zihe" last="Rao">Zihe Rao</name><name sortKey="Xie, Qingqe" sort="Xie, Qingqe" uniqKey="Xie Q" first="Qingqe" last="Xie">Qingqe Xie</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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