Transient expression of fusion gene coding for the HPV-16 epitopes fused to the sequence of potyvirus coat protein using different means of inoculation of Nicotiana benthamiana and Brassica rapa, cv. Rapa plants

Hoffmeisterová H., Čeřovská N., Moravec T., Plchová H., Folwarczna J., Veleminsky J.

Keywords: Edible vaccine, epitopes expression, E7 oncogene, Human papillomavirus, L2 structural protein, plant virus-based vector, Potato virus A, Potyvirus-like particles
Abstract: We describe the effect of different means of inoculation of Nicotiana benthamiana plants on the amount of transiently expressed fusion gene coding for the Human papillomavirus type 16 (HPV-16) epitopes fused to the sequence of recombinant Potato virus A coat protein (ACP). Epitope derived from minor capsid protein L2 was expressed as an N-terminal fusion with ACP while an epitope derived from E7 oncoprotein was fused to its C-terminus. The 885 bp construct (named L2ACPE7) was cloned into Potato virus X (PVX) based vector pGR106. In our experiments we compared two different means of agrobacteriummediated infection in N. benthamiana plants: agroinfection using either hypodermic syringe or vacuum infiltration. Furthermore, we describe the possibility of L2ACPE7 expression in an edible plant B. rapa, cv. Rapa. The presence and expression of resulting L2ACPE7 in B. rapa infected plants were confirmed by Immuno-Capture Reverse Transcription PCR (IC RT PCR), Plate-Trapped Antigen ELISA (PTAELISA) and SDS-PAGE/Western blot analysis and compared with the expression in N. benthamiana.
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IEB authors: Noemi Čeřovská, Hana Hoffmeisterová, Tomáš Moravec, Helena Plchová