Athukorala, Ajani, Helbig, Karla J., Mcsharry, Brian P., Forwood, Jade K., and Sarker, Subir (2024) An optimised protocol for the expression and purification of adenovirus core protein VII. Journal of Virological Methods, 326. 114907.

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Abstract

Adenovirus protein VII (pVII) is a highly basic core protein, bearing resemblance to mammalian histones. Despite its diverse functions, a comprehensive understanding of its structural intricacies and the mechanisms underlying its functions remain elusive, primarily due to the complexity of producing a good amount of soluble pVII. This study aimed to optimise the expression and purification of recombinant pVII from four different adenoviruses with a simple vector construct. This study successfully determined the optimal conditions for efficiently purifying pVII across four adenovirus species, revealing the differential preference for bacterial expression systems. The One Shot BL21 Star (DE3) proved favourable over Rosetta 2 (DE3) pLysS with consistent levels of expression between IPTG-induced and auto-induction. We demonstrated that combining chemical and mechanical cell lysis is possible and highly effective. Other noteworthy benefits were observed in using RNase during sample processing. The addition of RNase has significantly improved the quality and quantity of the purified protein as confirmed by chromatographic and western blot analyses. These findings established a solid groundwork for pVII purification methodologies and carry the significant potential to assist in unveiling the core structure of pVII, its arrangement within the core, DNA condensation intricacies, and potential pathways for nuclear transport.

Item ID:	83927
Item Type:	Article (Research - C1)
ISSN:	1879-0984
Keywords:	Adenovirus core protein, Core protein VII purification, DNA binding protein, pVII, RNase
Copyright Information:	© 2024 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Funders:	Australian Research Council (ARC)
Projects and Grants:	ARC DE200100367
Date Deposited:	29 Oct 2024 05:28
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