Castiglia, Daniela (2011) Glucosio-6P deidrogenasi citosolica di orzo: overespressione, caratterizzazione e regolazione da stress biotico e da sugar sensing. [Tesi di dottorato] (Unpublished)
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Item Type: | Tesi di dottorato |
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Resource language: | Italiano |
Title: | Glucosio-6P deidrogenasi citosolica di orzo: overespressione, caratterizzazione e regolazione da stress biotico e da sugar sensing |
Creators: | Creators Email Castiglia, Daniela daniela.castiglia@virgilio.it |
Date: | 30 November 2011 |
Number of Pages: | 148 |
Institution: | Università degli Studi di Napoli Federico II |
Department: | Biologia strutturale e funzionale |
Scuola di dottorato: | Scienze biologiche |
Dottorato: | Biologia applicata |
Ciclo di dottorato: | 24 |
Coordinatore del Corso di dottorato: | nome email Ricca, Ezio ericca@unina.it |
Tutor: | nome email Esposito, Sergio sergio.esposito@unina.it |
Date: | 30 November 2011 |
Number of Pages: | 148 |
Keywords: | Blumeria gramins,G6PDH, sugar sensing |
Settori scientifico-disciplinari del MIUR: | Area 05 - Scienze biologiche > BIO/04 - Fisiologia vegetale |
Date Deposited: | 06 Dec 2011 09:49 |
Last Modified: | 17 Jun 2014 06:04 |
URI: | http://www.fedoa.unina.it/id/eprint/8772 |
Collection description
A Glucose-6 phosphate dehydrogenase (G6PDH) encoding sequence was obtained from barley (Hordeum vulgare cv. Nure) roots and leaves. This sequence was 1767 bp long, and encoded for a 509 aminoacids long protein. The derived amino acid sequence was compared with known G6PDH sequences from various sources. The phylogenetic analisys suggests that this sequence would produce a cytosolic G6PDH isoform (cyt- G6PDH). This coding sequence was cloned in the expression vector pET-3d and recombinant cyt-G6PDH protein overexpressed in E.coli BL21 (DE3). The recombinant protein was purified and characterized. The enzyme specifically reacts vs potato cyt-G6PDH antibodies and exhibits kinetic parameters in agreement with those measured on cyt-G6PDH previously purified from barley roots. In vivo, the cyt-G6PDH expression level increased in the leaves upon infection with Blumeria graminis, more intensively in resistant plants with respect to susceptible plants, suggesting a main role of this isoform in the defence mechanisms developed in plants in both in specific and aspecific response to pathogens. Moreover, the regulation of cyt-G6PDH appears to be triggered by sugar sensing mechanisms: enzyme activity increased in hydroponic-grown plants upon 50mM fructose supply, sharper in roots than in leaves.
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