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[A preliminary study of the structure prediction and expression of SARS-CoV spike protein].

Identifieur interne : 004554 ( Main/Exploration ); précédent : 004553; suivant : 004555

[A preliminary study of the structure prediction and expression of SARS-CoV spike protein].

Auteurs : Mei-Ling Sun [République populaire de Chine] ; Jin-Hua Piao ; Chang-Cheng Yin ; Gong-She Yang ; Hua-Liang Huang

Source :

RBID : pubmed:16120590

Descripteurs français

English descriptors

Abstract

In this study, the encoding sequences of SARS-CoV spike protein were analyzed by bioinformatics methods, the structural characteristics and functions were forecasted based on available data. It suggests that the fragment of spike glycoprotein (S401-659) may be crucial for viral attachment and may be a major immunodominant epitope. Then the fragment was amplified and subcloned into expression vector pET28a(+) and pPIC9K. These two plasmids pET28a(+)-S and pPIC9K-S were transformed to E.coli strain BL21(DE3)-star and Pichia pastoris, respectively. SDS-PAGE and Western blot analysis showed that the recombinant protein was successfully expressed. The denatured inclusion bodies were purified with Ni-NTA chelate agarose and its purity can reach 90%.

PubMed: 16120590


Affiliations:


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Le document en format XML

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<term>Escherichia coli (genetics)</term>
<term>Gene Expression Regulation, Viral</term>
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<term>Membrane Glycoproteins (chemistry)</term>
<term>Membrane Glycoproteins (genetics)</term>
<term>Membrane Glycoproteins (metabolism)</term>
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<div type="abstract" xml:lang="en">In this study, the encoding sequences of SARS-CoV spike protein were analyzed by bioinformatics methods, the structural characteristics and functions were forecasted based on available data. It suggests that the fragment of spike glycoprotein (S401-659) may be crucial for viral attachment and may be a major immunodominant epitope. Then the fragment was amplified and subcloned into expression vector pET28a(+) and pPIC9K. These two plasmids pET28a(+)-S and pPIC9K-S were transformed to E.coli strain BL21(DE3)-star and Pichia pastoris, respectively. SDS-PAGE and Western blot analysis showed that the recombinant protein was successfully expressed. The denatured inclusion bodies were purified with Ni-NTA chelate agarose and its purity can reach 90%.</div>
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