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Use of phage display for isolation and characterization of single-chain variable fragments against dihydroflavonol 4-reductase from Petunia hybrida.
De Jaeger, G; Buys, E; Eeckhout, D; Bruyns, A M; De Neve, M; De Wilde, C; Gerats, T; Van Montagu, M; Fischer, R; Depicker, A.
Afiliación
  • De Jaeger G; Departement Genetica, Vlaams Interuniversitair Instituut voor Biotechnologie (VIB), Universiteit Gent, Belgium.
FEBS Lett ; 403(2): 116-22, 1997 Feb 17.
Article en En | MEDLINE | ID: mdl-9042949
To isolate specific single-chain variable (scFv) fragments against dihydroflavonol 4-reductase (DFR) from Petunia hybrida the phage display technology was used. DFR was overproduced in Escherichia coli, purified and used for immunization. From DFR-immunized mice, a phage display library was made starting from spleen mRNA using an optimized set of primers for V(H) and V(L) amplification. Several rounds of panning against recombinant DFR yielded five different scFv fragments, confirmed by subsequent DNA sequencing. They all specifically bound to recombinant DFR in ELISA and DFR in flower extracts on Western blot. These results show that phage display is a promising technology in plant molecular biology to obtain specific recombinant antibodies not only for ELISA and Western blot but also for in vivo applications in the long run.
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Plantas / Oxidorreductasas de Alcohol Límite: Animals Idioma: En Revista: FEBS Lett Año: 1997 Tipo del documento: Article País de afiliación: Bélgica Pais de publicación: Reino Unido
Buscar en Google
Añadir a Mi BVS
Imprimir
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PubMed Links

Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Plantas / Oxidorreductasas de Alcohol Límite: Animals Idioma: En Revista: FEBS Lett Año: 1997 Tipo del documento: Article País de afiliación: Bélgica Pais de publicación: Reino Unido