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Utilisation of the budding yeast Saccharomyces cerevisiae for the generation and isolation of non-lethal ricin A chain variants
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UNSPECIFIED (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. ISSN 0749-503X.
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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. Copyright (c) 2005 John Wiley & Sons, Ltd.
Item Type: | Journal Article | ||||
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Subjects: | Q Science > QD Chemistry T Technology > TP Chemical technology Q Science > QR Microbiology Q Science > QK Botany |
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Journal or Publication Title: | YEAST | ||||
Publisher: | JOHN WILEY & SONS LTD | ||||
ISSN: | 0749-503X | ||||
Official Date: | December 2005 | ||||
Dates: |
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Volume: | 22 | ||||
Number: | 16 | ||||
Number of Pages: | 11 | ||||
Page Range: | pp. 1287-1297 | ||||
Publication Status: | Published |
Data sourced from Thomson Reuters' Web of Knowledge
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