Utilisation of the budding yeast Saccharomyces cerevisiae for the generation and isolation of non-lethal ricin A chain variants
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 0749-503XFull text not available from this repository.
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|
|Number of Pages:||11|
|Page Range:||pp. 1287-1297|
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