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Provedor de dados:  Inra
País:  France
Título:  Single-chain variable fragments antibody specific toCorynespora cassiicola toxin, cassiicolin, reduces necrotic lesion formation in Hevea brasiliensis
Autores:  Sunderasan, E.
Kadir, R.A.
Pujade-Renaud, V.
de Lamotte, F.
Yeang, H.Y.
Nathan, S.
Data:  2009
Ano:  2009
Palavras-chave:  CLONE
Resumo:  Corynespora leaf disease poses a serious threat to rubber cultivation because infected leaves develop necrotic lesions and abscise, leaving the tree unproductive. The destructiveness of Corynespora cassiicola has been largely attributed to cassiicolin, a protein toxin secreted by the fungus. Recombinant antibody technology offers hope to curtail the disease whereby single-chain variable fragments (scFv) specific to cassiicolin could bind and deactivate the toxin in genetically modified rubber trees that harbour the antibody gene. A scFv phage library was constructed from heavy and light variable chains of IgG from cassiicolin immunized Balb/C mice spleen. Biopanning of the phage library yielded a scFv clone with high specificity to cassiicolin. The nucleotide sequence and deduced amino acid sequence information of the scFv were obtained. Hemagglutinin (HA)-tagged scFv expressed in Escherichia coli is discerned as a band at ca. 30 kDa on SDS-PAGE, and the corresponding band was detected by anti-HA IgG on a Western immunoblot. Deactivation of cassiicolin by the affinity-purified scFv was demonstrated in a detached-leaf bio-assay on selected susceptible Hevea clones (PB 260, RRIM 2020, RRIM 901 and RRIM 929). The assay was also performed on clones that are relatively more resistant to the fungus (RRIM 600 and GT-1), and the results were as expected. Thus, we have successfully demonstrated that the cassicolin-specific scFv can effectively reduce cassicolin toxicity
Tipo:  Journal Article
Idioma:  Inglês
Fonte:  Journal of General Plant Pathology. 2009, 15 (1) : 19-26

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