[The cloning, expression and structural analysis of putative unknown protein Orf 9b in SARS-CoV].
Identifieur interne : 001F99 ( Ncbi/Curation ); précédent : 001F98; suivant : 002000[The cloning, expression and structural analysis of putative unknown protein Orf 9b in SARS-CoV].
Auteurs : Wen Li [République populaire de Chine] ; Yang Liu ; Lei Lan ; Ming-Nan LiuSource :
- Bing du xue bao = Chinese journal of virology [ 1000-8721 ] ; 2009.
Descripteurs français
- KwdFr :
- Anticorps antiviraux (immunologie), Cadres ouverts de lecture, Clonage moléculaire, Expression des gènes, Humains, Protéines virales (), Protéines virales (génétique), Protéines virales (immunologie), Structure secondaire des protéines, Syndrome respiratoire aigu sévère (immunologie), Syndrome respiratoire aigu sévère (virologie), Virus du SRAS (), Virus du SRAS (génétique), Virus du SRAS (immunologie).
- MESH :
- génétique : Protéines virales, Virus du SRAS.
- immunologie : Anticorps antiviraux, Protéines virales, Syndrome respiratoire aigu sévère, Virus du SRAS.
- virologie : Syndrome respiratoire aigu sévère.
- Cadres ouverts de lecture, Clonage moléculaire, Expression des gènes, Humains, Protéines virales, Structure secondaire des protéines, Virus du SRAS.
English descriptors
- KwdEn :
- Antibodies, Viral (immunology), Cloning, Molecular, Gene Expression, Humans, Open Reading Frames, Protein Structure, Secondary, SARS Virus (chemistry), SARS Virus (genetics), SARS Virus (immunology), Severe Acute Respiratory Syndrome (immunology), Severe Acute Respiratory Syndrome (virology), Viral Proteins (chemistry), Viral Proteins (genetics), Viral Proteins (immunology).
- MESH :
- chemical , chemistry : Viral Proteins.
- chemical , genetics : Viral Proteins.
- chemical , immunology : Antibodies, Viral, Viral Proteins.
- chemistry : SARS Virus.
- genetics : SARS Virus.
- immunology : SARS Virus, Severe Acute Respiratory Syndrome.
- virology : Severe Acute Respiratory Syndrome.
- Cloning, Molecular, Gene Expression, Humans, Open Reading Frames, Protein Structure, Secondary.
Abstract
Orf 9b was amplified by PCR from SARS-CoV genome and cloned into the Nco I and Bam HI sites of the pET32c expression vector, and then recombinant plasmid pET32c-Orf 9b was constructed. The recombinant fusion protein Orf 9b was expressed by IPTG induction and purifed. After being cleaved by rEK, Orf 9b protein with MW 11 kD was separated and collected. It was demonstrated by ELISA that the purified Orf 9b protein could react with sera of SARS rehabilitaion patients but not with sera from healthy donors. CD and Infrared spectroscopy were used to predict the secondary structure of the purified Orf 9b protein. The distribution percentages for the the secondary structures of alpha-helix, beta-sheet, and random coil in the Orf 9b protein estimated by CD were 12.5%, 40%, and 47.5%, respectively, while the same parameters estimated by Infrared spectroscopy were 13.7%, 47.5%, and 37.9%, respectively. The results obtained by the two methods were substantially identical and showed that the structure of the Orf 9b protein consisted mostly of beta-sheet, and comprised only few alpha-helix. The acquisition of purified protein and the structural information presented herein may provide foundation for further functional study.
PubMed: 19769160
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SARS-CoV].</title><author><name sortKey="Li, Wen" sort="Li, Wen" uniqKey="Li W" first="Wen" last="Li">Wen Li</name><affiliation wicri:level="1"><nlm:affiliation>Department of Cell Biology, School of Basic Medical Science, Capital Medical University, Beijing 100069, China. liwen@ccmu.edu.cn</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Cell Biology, School of Basic Medical Science, Capital Medical University, Beijing 100069</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Liu, Yang" sort="Liu, Yang" uniqKey="Liu Y" first="Yang" last="Liu">Yang Liu</name></author><author><name sortKey="Lan, Lei" sort="Lan, Lei" uniqKey="Lan L" first="Lei" last="Lan">Lei Lan</name></author><author><name sortKey="Liu, Ming Nan" sort="Liu, Ming Nan" uniqKey="Liu M" first="Ming-Nan" last="Liu">Ming-Nan Liu</name></author></analytic><series><title level="j">Bing du xue bao = Chinese journal of virology</title><idno type="ISSN">1000-8721</idno><imprint><date when="2009" type="published">2009</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antibodies, Viral (immunology)</term><term>Cloning, Molecular</term><term>Gene Expression</term><term>Humans</term><term>Open Reading Frames</term><term>Protein Structure, Secondary</term><term>SARS Virus (chemistry)</term><term>SARS Virus (genetics)</term><term>SARS Virus (immunology)</term><term>Severe Acute Respiratory Syndrome (immunology)</term><term>Severe Acute Respiratory Syndrome (virology)</term><term>Viral Proteins (chemistry)</term><term>Viral Proteins (genetics)</term><term>Viral Proteins (immunology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Anticorps antiviraux (immunologie)</term><term>Cadres ouverts de lecture</term><term>Clonage moléculaire</term><term>Expression des gènes</term><term>Humains</term><term>Protéines virales ()</term><term>Protéines virales (génétique)</term><term>Protéines virales (immunologie)</term><term>Structure secondaire des protéines</term><term>Syndrome respiratoire aigu sévère (immunologie)</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Virus du SRAS ()</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (immunologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Viral Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Viral Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Viral</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protéines virales</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps antiviraux</term><term>Protéines virales</term><term>Syndrome respiratoire aigu sévère</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</term><term>Severe Acute Respiratory Syndrome</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>Cloning, Molecular</term><term>Gene Expression</term><term>Humans</term><term>Open Reading Frames</term><term>Protein Structure, Secondary</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Cadres ouverts de lecture</term><term>Clonage moléculaire</term><term>Expression des gènes</term><term>Humains</term><term>Protéines virales</term><term>Structure secondaire des protéines</term><term>Virus du SRAS</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Orf 9b was amplified by PCR from SARS-CoV genome and cloned into the Nco I and Bam HI sites of the pET32c expression vector, and then recombinant plasmid pET32c-Orf 9b was constructed. The recombinant fusion protein Orf 9b was expressed by IPTG induction and purifed. After being cleaved by rEK, Orf 9b protein with MW 11 kD was separated and collected. It was demonstrated by ELISA that the purified Orf 9b protein could react with sera of SARS rehabilitaion patients but not with sera from healthy donors. CD and Infrared spectroscopy were used to predict the secondary structure of the purified Orf 9b protein. The distribution percentages for the the secondary structures of alpha-helix, beta-sheet, and random coil in the Orf 9b protein estimated by CD were 12.5%, 40%, and 47.5%, respectively, while the same parameters estimated by Infrared spectroscopy were 13.7%, 47.5%, and 37.9%, respectively. The results obtained by the two methods were substantially identical and showed that the structure of the Orf 9b protein consisted mostly of beta-sheet, and comprised only few alpha-helix. The acquisition of purified protein and the structural information presented herein may provide foundation for further functional study.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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