Polyclonal antibodies to a recombinant coat protein of Potato virus A

Specific mouse antibodies against a recombinant coat protein (CP) of Potato virus A (PVA) were produced. The PVA CP gene was cloned in an expression vector pMPM4omega. After expression in Escherichia coli the presence of the expressed CP was proved by Western blot analysis using polyclonal and monoc...

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Bibliographic Details
Published inActa virologica (Anglickâa verze) Vol. 46; no. 3; p. 147
Main Authors Cerovská, N, Moravec, T, Velemínský, J
Format Journal Article
LanguageEnglish
Published Slovakia 2002
Subjects
Antibodies, Monoclonal - immunology
Antibodies, Viral - analysis
Antibodies, Viral - immunology
Blotting, Western
Capsid Proteins - genetics
Capsid Proteins - immunology
Cloning, Molecular
Enzyme-Linked Immunosorbent Assay
Escherichia coli - metabolism
Microscopy, Electron
Potyvirus - genetics
Potyvirus - immunology
Recombinant Proteins - chemistry
Recombinant Proteins - immunology
Solanum tuberosum - virology
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Summary:Specific mouse antibodies against a recombinant coat protein (CP) of Potato virus A (PVA) were produced. The PVA CP gene was cloned in an expression vector pMPM4omega. After expression in Escherichia coli the presence of the expressed CP was proved by Western blot analysis using polyclonal and monoclonal antibodies (MAbs). The expressed CP was purified by centrifugation in CsCl density gradient or on a sucrose cushion. The production of virus-like particles (VLPs) was proved by electron microscopy. The purified CP was used for preparation of a mouse antiserum which had a titer of 1:1024 in ELISA and reacted specifically in Western blot analysis and indirect plate-trapped antigen ELISA (PTA-ELISA).
ISSN:0001-723X