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Identifier |
000443041 |
Title |
Protein biotechnology-based approaches for the functional and structural characterization of insect enzymes |
Alternative Title |
Μέθοδοι πρωτεϊνικής βιοτεχνολογίας και στον λειτουργικό και δομικό χαρακτηρισμό πρωτεϊνών εντόμων |
Author
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Σαράφογλου, Χαρά Δ.
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Thesis advisor
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Βόντας, Ιωάννης
Μπαλαμπανίδου, Βασιλεία
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Reviewer
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Κοκκινίδης, Μιχαήλ
Παυλίδης, Ιωάννης
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Abstract |
Since the practice of agriculture first begun, the loss of yield due to several insect pests
was apparent. Even in recent years, with advances in agricultural sciences, losses due to pests
and diseases range from 10-90%. Not limited to agricultural damage, insects are also
responsible for disease transmission having a profound impact on human and livestock health.
To address economy and health issues caused by these insects, various field techniques are
used for insect control that can be grouped into three categories: mechanical, biological, and
chemical.
Chemical methods are related to the use of insecticides targeting harmful insect
species with the intention of eliminating them. Since the first applications of insecticides,
insects, pressured by natural selection, started developing resistance mechanisms against
them. This created the need of new methods of action in the form of new chemicals or new
targets, forming this never-ending cycle.
Protein biotechnology-based approaches is a powerful tool used to win the race
against insecticide resistance. In vitro techniques are used to isolate and characterize existing
or novel insecticide targets. Furthermore, resistance mechanisms can be studied in order to
identify patterns and eventually resolve problems by slightly altering existing commercially
available insecticides or even by designing new chemicals that by-pass insect defenses. For
the purpose of this work, two such examples are being examined.
As a new promising target, type III geranylgeranyl diphosphate synthase from
Helicoverpa armigera was isolated and examined. After protein purification, an assay was
adapted to measure activity and determine kinetic values. Through homology modeling,
substrate binding patterns were evaluated, and new chemicals were designed using in silico
techniques. These chemicals were later tested in the assay to determine potency by highthroughput screening.
Finally, Cytochrome P450 from Bemicia tabaci, an enzyme with detoxification
function, was studied as it is implicated in insecticide resistance. In this case, the main
objective was to establish a reproducible protocol of bacterial expression and purification of
the enzyme while keeping it functional in lab conditions. Due to this enzyme’s properties the
above proved to be challenging and involved protein engineering strategies to improve
efficiency.
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Language |
English |
Subject |
Geranylgeranyl diphosphate synthase |
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Insecticide targets |
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Pest control |
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Resistance mechanism |
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Αντιμετώπιση εντόμων |
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Μηχανισμοί ανθεκτικότητας |
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Μοριακή εντομολογία |
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Στόχοι εντομοκτόμων |
Issue date |
2021-11-26 |
Collection
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School/Department--School of Sciences and Engineering--Department of Biology--Post-graduate theses
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Type of Work--Post-graduate theses
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Permanent Link |
https://elocus.lib.uoc.gr//dlib/f/0/2/metadata-dlib-1634740404-298280-28316.tkl
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Views |
361 |