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
Subjects:	Q Science > QD Chemistry T Technology > TP Chemical technology Q Science > QR Microbiology Q Science > QK Botany
Journal or Publication Title:	YEAST
Publisher:	JOHN WILEY & SONS LTD
ISSN:	0749-503X
Date:	December 2005
Volume:	22
Number:	16
Number of Pages:	11
Page Range:	pp. 1287-1297
Publication Status:	Published
URI:	http://wrap.warwick.ac.uk/id/eprint/33976

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