Gallo, Giovanni (2020) Development of biosensors for monitoring of envirnmental pollutants. [Tesi di dottorato]


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Item Type: Tesi di dottorato
Resource language: English
Title: Development of biosensors for monitoring of envirnmental pollutants
Date: 10 March 2020
Number of Pages: 159
Institution: Università degli Studi di Napoli Federico II
Department: Biologia
Dottorato: Biotecnologie
Ciclo di dottorato: 32
Coordinatore del Corso di dottorato:
Bartolucci, SimonettaUNSPECIFIED
Fiorentino, GabriellaUNSPECIFIED
Date: 10 March 2020
Number of Pages: 159
Keywords: biosensor, heavy metals resistance system, thermophiles
Settori scientifico-disciplinari del MIUR: Area 05 - Scienze biologiche > BIO/10 - Biochimica
Date Deposited: 06 Apr 2020 09:26
Last Modified: 10 Nov 2021 10:22

Collection description

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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