The expression of SARS-CoV M gene in P. Pastoris and the diagnostic utility of the expression product
Identifieur interne : 005A53 ( Main/Exploration ); précédent : 005A52; suivant : 005A54The 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.
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- Pascal (Inist)
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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 M gene fragment was amplified and cloned it into the Pichia Pastoris 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.
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Pastoris and the diagnostic utility of the expression product</title><author><name sortKey="Xueqing Han" sort="Xueqing Han" uniqKey="Xueqing Han" last="Xueqing Han">XUEQING HAN</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Laboratory of Structural Biology, Tsinghua University and National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Science</s1><s2>Beijing</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>République populaire de Chine</country><placeName><settlement type="city">Pékin</settlement></placeName></affiliation><affiliation wicri:level="3"><inist:fA14 i1="02"><s1>Lanzhou Institute of Veterinary Medicine, Chinese Academy of Agricultural Sciences, Baishiquio Road 30</s1><s2>Beijing</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>République populaire de Chine</country><placeName><settlement 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Science</s1><s2>Beijing</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>République populaire de Chine</country><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Xiangtao Liu" sort="Xiangtao Liu" uniqKey="Xiangtao Liu" last="Xiangtao Liu">XIANGTAO LIU</name><affiliation wicri:level="3"><inist:fA14 i1="02"><s1>Lanzhou Institute of Veterinary Medicine, Chinese Academy of Agricultural Sciences, Baishiquio Road 30</s1><s2>Beijing</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>République populaire de Chine</country><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Xiaojing He" sort="Xiaojing He" uniqKey="Xiaojing He" last="Xiaojing He">XIAOJING HE</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Laboratory of Structural Biology, Tsinghua University and National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Science</s1><s2>Beijing</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>République populaire de Chine</country><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Xuepeng Cai" sort="Xuepeng Cai" uniqKey="Xuepeng Cai" last="Xuepeng Cai">XUEPENG CAI</name><affiliation wicri:level="3"><inist:fA14 i1="02"><s1>Lanzhou Institute of Veterinary Medicine, Chinese Academy of Agricultural Sciences, Baishiquio Road 30</s1><s2>Beijing</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>République populaire de Chine</country><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Qingqe Xie" sort="Qingqe Xie" uniqKey="Qingqe Xie" last="Qingqe Xie">QINGQE XIE</name><affiliation wicri:level="3"><inist:fA14 i1="02"><s1>Lanzhou Institute of Veterinary Medicine, Chinese Academy of Agricultural Sciences, Baishiquio Road 30</s1><s2>Beijing</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>République populaire de Chine</country><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Zihe Rao" sort="Zihe Rao" uniqKey="Zihe Rao" last="Zihe Rao">ZIHE RAO</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Laboratory of Structural Biology, Tsinghua University and National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Science</s1><s2>Beijing</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>5 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>République populaire de Chine</country><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author></analytic><series><title level="j" type="main">Journal of virological methods</title><title level="j" type="abbreviated">J. virol. methods</title><idno type="ISSN">0166-0934</idno><imprint><date when="2004">2004</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of virological methods</title><title level="j" type="abbreviated">J. virol. methods</title><idno type="ISSN">0166-0934</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antigenicity</term><term>Gene expression</term><term>Method</term><term>Microbiology</term><term>Recombinant protein</term><term>Severe acute respiratory syndrome</term><term>Severe acute respiratory syndrome virus</term><term>Virology</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Expression génique</term><term>Protéine recombinante</term><term>Antigénicité</term><term>Microbiologie</term><term>Méthode</term><term>Virologie</term><term>Syndrome respiratoire aigu sévère</term><term>Virus syndrome respiratoire aigu sévère</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">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 M gene fragment was amplified and cloned it into the Pichia Pastoris 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.</div></front></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="Xueqing Han" sort="Xueqing Han" uniqKey="Xueqing Han" last="Xueqing Han">XUEQING HAN</name></noRegion><name sortKey="Bartlam, Mark" sort="Bartlam, Mark" uniqKey="Bartlam M" first="Mark" last="Bartlam">Mark Bartlam</name><name sortKey="Jin, Ying Hua" sort="Jin, Ying Hua" uniqKey="Jin Y" first="Ying-Hua" last="Jin">Ying-Hua Jin</name><name sortKey="Qingqe Xie" sort="Qingqe Xie" uniqKey="Qingqe Xie" last="Qingqe Xie">QINGQE XIE</name><name sortKey="Xiangtao Liu" sort="Xiangtao Liu" uniqKey="Xiangtao Liu" last="Xiangtao Liu">XIANGTAO LIU</name><name sortKey="Xiaojing He" sort="Xiaojing He" uniqKey="Xiaojing He" last="Xiaojing He">XIAOJING HE</name><name sortKey="Xuepeng Cai" sort="Xuepeng Cai" uniqKey="Xuepeng Cai" last="Xuepeng Cai">XUEPENG CAI</name><name sortKey="Xueqing Han" sort="Xueqing Han" uniqKey="Xueqing Han" last="Xueqing Han">XUEQING HAN</name><name sortKey="Zihe Rao" sort="Zihe Rao" uniqKey="Zihe Rao" last="Zihe Rao">ZIHE RAO</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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