Allen, S. C. H., Byron, A., Lord, J. M., Davey, J., Roberts, L. M. and Ladds, G. (2005) Utilisation of the budding yeast Saccharomyces cerevisiae for the generation and isolation of non-lethal ricin A chain variants. Yeast. 22(16), pp. 1287-1297. 0749-503X.
| Item Type: | Article |
|---|---|
| Abstract: | Knowledge of the uptake, membrane translocation, refolding and ribosome interaction of the ribosome-inactivating toxin ricin is incomplete at the present time. Ricin A chain (RTA) is the catalytic subunit of holotoxin and is also of particular interest as a vaccine candidate. For many studies into the uptake and immunological applications of ricin, it is essential to have inactive variants. Here, following error-prone polymerase chain reaction of the RTA open reading frame, we have used a modified gap-repair protocol in Saccharomyces cerevisiae to show that it is possible to rapidly generate a panel of inactive RTA mutants. Since yeast cells have ribosomes that are highly sensitive to RTA, we utilized a genetic selection based on the viability of transformants. This enabled the recovery of a number of mutations, some not previously identified, which permitted production of full-length but non-toxic RTA proteins. Such disarmed toxins may have utility as tools to study the cytosolic entry and action of RTA, and as potential vaccine candidates |
| Uncontrolled Keywords: | gap repair, yeast, mutation, ricin A chain |
| Subjects: | Q Science > QR Microbiology > QR151 Yeasts |
| Creators: | Allen, Stuart C H, Byron, A, Lord, J M, Davey, J, Roberts, L M and Ladds, G |
| Publisher: | John Wiley & Sons |
| Faculties, Divisions and Institutes: |
University Faculties, Divisions and Research Centres - OLD > Faculty of Health & Society > Sports, Exercise & Life Sciences Faculties > Faculty of Health & Society > Sports, Exercise & Life Sciences |
| Date: | December 2005 |
| Date Type: | Publication |
| Page Range: | pp. 1287-1297 |
| Journal or Publication Title: | Yeast |
| Volume: | 22 |
| Number: | 16 |
| Language: | English |
| DOI: | https://doi.org/10.1002/yea.1330 |
| ISSN: | 0749-503X |
| Status: | Published / Disseminated |
| URI: | http://nectar.northampton.ac.uk/id/eprint/5328 |
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