The expression of SARS-CoV M gene in P. Pastoris and the diagnostic utility of the expression product
Identifieur interne : 005A53 ( Main/Curation ); 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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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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<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>
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<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>
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<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>
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