The aim of the thesis project was to verify the involvement of CYP3A28 in the metabolism of aflatoxin B1 (AFB1), in a bovine foetus liver cell line (BFH12), being this enzyme considered as one of the main bioactivators of mycotoxin. BFH12 was subjected to a gene deletion method, CRISPR-Cas9, through which it was possible to obtain a cell clone deleting CYP3A28. On this clone, tests were performed to confirm and verify the involvement of CYP3A28 in the metabolism of AFB1, such as: cytotoxicity, catalytic activity, Western-blot and Real-Time PCR.
Lo scopo del progetto di tesi è stato quello di verificare il coinvolgimento del CYP3A28 nel metabolismo dell’aflatossina B1 (AFB1), in una linea cellulare di fegato di feto bovino (BFH12), essendo tale enzima considerato come uno dei principali bioattivatori della micotossina. BFH12 è stata sottoposta ad una metodica di delezione genica, CRISPR-Cas9, attraverso cui è stato possibile ottenere un clone cellulare deleto per il CYP3A28. Su tale clone sono stati eseguiti dei test di conferma e di verifica del coinvolgimento del CYP3A28 nel metabolismo di AFB1, quali: citotossicità, attività catalitica, Western-blot e Real-Time PCR.
Definizione del ruolo del CYP3A28 nell'aflatossicosi bovina mediante applicazione della metodica CRISPR-Cas9
PALANCA, MARTINA
2022/2023
Abstract
The aim of the thesis project was to verify the involvement of CYP3A28 in the metabolism of aflatoxin B1 (AFB1), in a bovine foetus liver cell line (BFH12), being this enzyme considered as one of the main bioactivators of mycotoxin. BFH12 was subjected to a gene deletion method, CRISPR-Cas9, through which it was possible to obtain a cell clone deleting CYP3A28. On this clone, tests were performed to confirm and verify the involvement of CYP3A28 in the metabolism of AFB1, such as: cytotoxicity, catalytic activity, Western-blot and Real-Time PCR.File | Dimensione | Formato | |
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https://hdl.handle.net/20.500.12608/52170