Gallo, Giovanni
(2020)
Development of biosensors for monitoring of envirnmental pollutants.
[Tesi di dottorato]
| Tipologia del documento: |
Tesi di dottorato
|
| Lingua: |
English |
| Titolo: |
Development of biosensors for monitoring of envirnmental pollutants |
| Autori: |
| Autore | Email |
|---|
| Gallo, Giovanni | giovanni.gallo2@unina.it |
|
| Data: |
10 Marzo 2020 |
| Numero di pagine: |
159 |
| Istituzione: |
Università degli Studi di Napoli Federico II |
| Dipartimento: |
Biologia |
| Dottorato: |
Biotecnologie |
| Ciclo di dottorato: |
32 |
| Coordinatore del Corso di dottorato: |
| nome | email |
|---|
| Moracci, Marco | marco.moracci@unina.it |
|
| Tutor: |
| nome | email |
|---|
| Bartolucci, Simonetta | [non definito] | | Fiorentino, Gabriella | [non definito] |
|
| Data: |
10 Marzo 2020 |
| Numero di pagine: |
159 |
| Parole chiave: |
biosensor, heavy metals resistance system, thermophiles |
| Settori scientifico-disciplinari del MIUR: |
Area 05 - Scienze biologiche > BIO/10 - Biochimica |
[error in script]
[error in script]
| Depositato il: |
06 Apr 2020 09:26 |
| Ultima modifica: |
10 Nov 2021 10:22 |
| URI: |
http://www.fedoa.unina.it/id/eprint/13045 |
Abstract
The high concentrations of heavy metals are the cause of one of the most serious pollution problems of our time. Different chemical or physical methods have been developed to treat polluted sites, but the problem persists given the necessity to dispose the treated waste and any, probably even more toxic, chemical by-product; so, considering advances in biotechnology, there is interest in the development of biosensors for monitoring polluted areas as well as bioremediation techniques.
The aim of this PhD thesis was the development of whole-cell biosensors for monitoring of environmental pollutants starting from the characterization of the molecular mechanisms of heavy metal resistance in thermophilic microorganisms.
The first part of this PhD-thesis focuses on the characterization of the TtSmtB, a trans-acting repressor belonging to the ArsR/SmtB family, involved in the metal sensing and in transcriptional regulation.
The characterization of TtSmtB was carried out in order to investigate the molecular determinants of metal binding, DNA binding and to identify target promoter regions.
The second part of this PhD thesis was focused on the development of whole-cell biosensors using two different approaches: plasmid-based or genome-based detection systems.
The first whole-cell biosensor was obtained starting from the knowledge acquired on the biological function of TtSmtB, where on a single plasmid we can found the promoter recognized by TtSmtB and the reporter gene.
The second whole-cell biosensor was obtained by engineering Thermus thermophilus HB27 through a CRISPR-Cas9 system that allowed us to insert the reporter gene into the genome, under the control of a responsive promoter, allowing us to improve the sensitivity of our reporter system.
The last part of this project was focused on the characterization of a new strain of Geobacillus stearothermophilus was isolated from a hot spring located in a solfataric area of Pozzuoli (Napoli).
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