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Production of Polyclonal Antibodies to Potato virus X Using Recombinant Coat Protein
Author(s) -
Cerovska Noemi,
Moravec Tomas,
Plchova Helena,
Hoffmeisterova Hana,
Folwarczna Jitka,
Dedic Petr
Publication year - 2010
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/j.1439-0434.2009.01580.x
Subject(s) - potato virus x , polyclonal antibodies , microbiology and biotechnology , biology , recombinant dna , virology , antibody , potexvirus , reverse transcription polymerase chain reaction , escherichia coli , virus , plant virus , coat protein , gene , gene expression , rna , biochemistry , immunology
Polyclonal antibodies to recombinant Potato virus X (PVX) coat protein (PVX‐CP) were developed and their effectiveness determined in different ELISA protocols. The PVX‐CP gene was amplified by reverse transcription‐polymerase chain reaction, cloned and expressed in Escherichia coli . For immunization, the CP fractions from bacterial lysate were purified either by simple fractionation or by excision from sodium dodecyl sulphate gels. The PVX‐CP was injected into rabbits for antibody production. The PVX‐CP antibodies reacted in an indirect plate trapped antigen enzyme‐linked immunosorbent assay and immunoblot assay and were useful for the detection of a broad spectrum of isolates of PVX.