Issue when expressing a recombinant protein under the control of p 35S in Nicotiana tabacum BY-2 cells

Catherine Navarre; Rik Orval; Marie Peeters; Nicolas Bailly; François Chaumont

doi:10.3389/fpls.2023.1266775

Issue when expressing a recombinant protein under the control of p 35S in Nicotiana tabacum BY-2 cells

Front Plant Sci. 2023 Nov 13:14:1266775. doi: 10.3389/fpls.2023.1266775. eCollection 2023.

Authors

Catherine Navarre¹, Rik Orval¹, Marie Peeters¹, Nicolas Bailly¹, François Chaumont¹

Affiliation

¹ Louvain Institute of Biomolecular Science and Technology (LIBST), UCLouvain, Louvain-la-Neuve, Belgium.

Abstract

Several recombinant proteins have been successfully produced in plants. This usually requires Agrobacterium-mediated cell transformation to deliver the T-DNA into the nucleus of plant cells. However, some genetic instability may threaten the integrity of the expression cassette during its delivery via A. tumefaciens, especially when the protein of interest is toxic to the bacteria. In particular, we found that a Tn3 transposon can be transferred from the pAL4404 Ti plasmid of A. tumefaciens LBA4404 into the expression cassette when using the widely adopted 35S promoter, thereby damaging T-DNA and preventing correct expression of the gene of interest in Nicotiana tabacum BY-2 suspension cells.

Keywords: Agrobacterium; BY-2 cells; LBA4404; Ti plasmid; bacterial transposon.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by the SPW Wallonie-Belgium in WALInnov project “Glycocell” (1810010). Nicolas Bailly is a recipient of a fellowship from the Fonds pour la Formation à la Recherche dans l’Industrie et l’Agriculture (FRIA, Belgium). Marie Peeters is a recipient of a fellowship from the Fonds National de la Recherche Scientifique (FNRS, Belgium).