biostudies-literatureUnknown14Navarre CSeveral recombinant proteins have been successfully produced in plants. This usually requires <i>Agrobacterium</i>-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 <i>A. tumefaciens</i>, 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 <i>A. tumefaciens</i> LBA4404 into the expression cassette when using the widely adopted <i>35S</i> promoter, thereby damaging T-DNA and preventing correct expression of the gene of interest in <i>Nicotiana tabacum</i> BY-2 suspension cells.Frontiers in plant science1266775https://www.ebi.ac.uk/biostudies/studies/S-EPMC10679441biostudies-literatureIssue when expressing a recombinant protein under the control of p<i>35S</i> in <i>Nicotiana tabacum</i> BY-2 cells.PMC10679441Orval RBailly NNavarre CChaumont FPeeters MfalseIssue when expressing a recombinant protein under the control of p<i>35S</i> in <i>Nicotiana tabacum</i> BY-2 cells.Several recombinant proteins have been successfully produced in plants. This usually requires <i>Agrobacterium</i>-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 <i>A. tumefaciens</i>, 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 <i>A. tumefaciens</i> LBA4404 into the expression cassette when using the widely adopted <i>35S</i> promoter, thereby damaging T-DNA and preventing correct expression of the gene of interest in <i>Nicotiana tabacum</i> BY-2 suspension cells.2023-01-01T00:00:00Z20232026-05-29T00:18:57.944Z2025-02-19T00:38:53.431ZS-EPMC106794413802388110.3389/fpls.2023.1266775