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Lookup NU author(s): Professor Neil Boonham,
Professor Rick Mumford
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Partial recombinant secA proteins were produced from six different phytoplasma isolates representing five 16Sr groups and the expressed, purified recombinant (partial secA) protein from Cape St. Paul wilt disease phytoplasma (CSPWD, 16SrXXII) was used to immunise mice. Monoclonal antibodies (mAbs) were selected by screening hybridoma supernatants for binding to the recombinant proteins. To characterise the binding to proteins from different phytoplasmas, the antibodies were screened by ELISA and western blotting, and epitope mapping was undertaken. Eight different mAbs with varying degrees of specificity against recombinant proteins from different phytoplasma groups were selected. Western blotting revealed that the mAbs bind to proteins in infected plant material, two of which were specific for phytoplasmas. ELISA testing of infected material, however, gave negative results suggesting that either secA was not expressed at sufficiently high levels, or conformational changes of the reagents adversely affected detection. This work has shown that the phytoplasma secA gene is not a suitable antibody target for routine detection, but has illustrated proof of principle for the methodology. © 2014 Crown Copyright.
Author(s): Hodgetts J, Johnson G, Perkins K, Ostoja-Starzewska S, Boonham N, Mumford R, Dickinson M
Publication type: Article
Publication status: Published
Journal: Molecular Biotechnology
Print publication date: 01/09/2014
Online publication date: 21/05/2014
Acceptance date: 01/01/1900
ISSN (print): 1073-6085
ISSN (electronic): 1559-0305
Publisher: Humana Press Inc.
PubMed id: 24845751
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