Expression of a Sheep Pox Virus Gene in Plant Systems under the Control of Plant Viral Regulatory Elements and with Sub-Cellular Targeting

DaniyarBeisenov*, GulshanStanbekova, Leila Nadirova, Andrey Zhigailov, BulatIskakov Corresponding Authors's Email: beisenov.d@gmail.com

Aitkhozhin Institute of Molecular Biology and Biochemistry, 86, Dosmukhamedov Str., Almaty, Kazakhstan

DOI : http://dx.doi.org/10.13005/bbra/1995

ABSTRACT: The aim of this work was to investigate the feasibility of producing the sheep pox virus surface antigen that is an ortholog of vaccinia viral A27L in wheat germ cell free system and then in transgenic tobacco plants. Insertion into the A27L-mRNA 5ʹ-untranslated region (5ʹ‑UTR) of various translational enhancers (TEs) from plant viruses, as well as an artificial TE, allowed to increase the level of A27L protein synthesis in a wheat germ cell-free system. When the 5ʹ‑UTR of potato virus Y genomic RNA was used as a TE, synthesis of an additional polypeptide that was approximately 2 kDa larger than the primary product of translation was observed in both plant and mammalian cell-free systems. The DNA constructs used for stable transformation of tobacco plants, besides 5ʹ‑TEs, contained the transit-peptide-coding sequences for targeted accumulation of synthesized A27L protein. The amount of recombinant protein in the leaves of transgenic plants varied from 0.01 to 0.03 % of the total soluble protein.

KEYWORDS: A27L protein, Potato virus Y Sheep pox virus,

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