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Identifier

000443710

Title

Functional expression and characterization of cytochrome P450s, associated with pesticide toxicity

Alternative Title

Λειτουργική έκφραση και χαρακτηρισμός κυτοχρωμικών μονοξειδάσων P450s που συνδέονται με την τοξικότητα που προκαλούν τα παρασιτοκτόνα

Author

Καϊλή, Σταυρούλα Ι.

Thesis advisor

Βόντας, Ιωάννης

Reviewer

Παυλίδης, Ιωάννης
Χαλεπάκης, Γεώργιος

Abstract

Ιnsecticide selectivity and resistance is an ongoing challenge concerning both crop pest control and insect-born diseases control, and it has been largely associated with cytochrome P450 monoοxyganases’ metabolism. Functional characterization of P450s is of great importance, in order to validate their involvement in insecticide resistance and to develop selective and safe insecticides (e.g. for bee pollinators) and insecticide synergists. In the present study, two distinct cases of P450s that have been associated with resistance to insecticides were heterologously expressed in E. coli for their functional characterization. Firstly, HaCYP6B7, which was previously found to be associated with fenvalerate metabolism in Helicoverpa armigera, was co-expressed in E. coli together with Musca domestica CPR (MdCPR). HaCYP6B7 was expressed in its native and truncated form in the membrane or in the cytoplasm of bacteria, respectively. The recombinant enzyme was assayed for its catalytic activity against model substrates. In the second part of this study, the aim was to heterologously express in E. coli BoCYP6G2 and BoCYP6A23 that have been related with resistance to α-cypermethrin in Bactrocera oleae. The two P450s were successfully cloned into expressing vectors and BoCYP6G2 proceeded to membrane targeting experiments together with Bactrocera oleae CPR (BoCPR).

Language

English

Subject

Heterologous expression in E.Coli

Ετερόλογη έκφραση στο σύστημα E.Coli

Κυτόχρωμα P450

Τοξικότητα παρασιτοκτόνων

Issue date

2021-11-26

Collection

School/Department--School of Sciences and Engineering--Department of Biology--Post-graduate theses

Type of Work--Post-graduate theses