Castiglia, Daniela
(2011)
Glucosio-6P deidrogenasi citosolica di orzo: overespressione, caratterizzazione e regolazione da stress biotico e da sugar sensing.
[Tesi di dottorato]
(Unpublished)
| Item Type: |
Tesi di dottorato
|
| Lingua: |
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 |
| Uncontrolled Keywords: |
Blumeria gramins,G6PDH, sugar sensing |
| Settori scientifico-disciplinari del MIUR: |
Area 05 - Scienze biologiche > BIO/04 - Fisiologia vegetale |
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| Date Deposited: |
06 Dec 2011 09:49 |
| Last Modified: |
17 Jun 2014 06:04 |
| URI: |
http://www.fedoa.unina.it/id/eprint/8772 |
Abstract
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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