1. Evaluation of different strategies to produce Vibrio cholerae ParE2 toxin.
- Author
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Girardin Y, Galle M, Vanden Abeele Y, De Greve H, and Loris R
- Subjects
- Operon, Bacterial Proteins genetics, Bacterial Proteins metabolism, Bacterial Toxins genetics, Antitoxins genetics, Antitoxins metabolism, Vibrio cholerae genetics
- Abstract
Toxin-antitoxin (TA) systems are small operons that are omnipresent in bacteria and archaea with suggested roles in stabilization of mobile genetic elements, bacteriophage protection, stress response and possibly persister formation. A major bottleneck in the study of TA toxins is the production of sufficient amounts of well-folded, functional protein. Here we examine alternative approaches for obtaining the VcParE2 toxin from Vibrio cholerae. VcParE2 can be successfully produced via bacterial expression in presence of its cognate antitoxin VcParD2, followed by on-column unfolding and refolding. Alternatively, the toxin can be expressed in Spodoptera frugiperda (Sf9) insect cells. The latter requires disruption of the VcparE2 gene via introduction of an insect cell intron. Both methods provide protein with similar structural and functional characteristics., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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