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ORIGINAL RESEARCH article
Front. Plant Sci.
Sec. Plant Biotechnology
Volume 16 - 2025 | doi: 10.3389/fpls.2025.1579465
Impact of Codon Optimization on vip3Aa11 Gene Expression and Insecticidal Efficacy in Maize
Provisionally accepted- Biotechnology Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Beijing, China
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Codon optimization is critical for high expression of foreign genes in heterologous systems. The vip3Aa11 gene from Bacillus thuringiensis is a promising candidate for controlling Spodoptera frugiperda. To develop insect-resistant maize, we designed two codon-optimized vip3Aa11 variants (vip3Aa11-m1 and vip3Aa11-m2) based on maize codon usage bias. Both recombinant proteins expressed in Escherichia coli exhibited high insecticidal activity. However, in transgenic maize, Vip3Aa11-m1 exhibited strong insecticidal activity against Spodoptera frugiperda and Spodoptera exigua, while Vip3Aa11-m2 lost activity despite identical amino acid sequences. RT-PCR analysis confirmed that both genes were transcribed correctly, but western blot results demonstrated a smaller product for vip3Aa11-m2, suggesting a translation-level alteration. Segment replacement and point mutation experiments in maize protoplasts demonstrated that the synonymous codon AAT (Asn) at the fourth amino acid position in vip3Aa11-m2 was associated with the production of a truncated protein, suggesting that the AAT codon may influence the selection of the translation initiation site, potentially shifting it to a downstream ATG (Met) codon. These findings not only reveal the critical role of codon context in translation initiation and protein integrity but also provide a novel strategy for optimizing foreign genes in crop improvement, particularly offering valuable insights for engineering insect-resistant maize using Bt genes.
Keywords: codon optimization, vip3Aa11 gene, transgenic maize, Spodoptera frugiperda, translation initiation, Synonymous codon substitution
Received: 19 Feb 2025; Accepted: 11 Apr 2025.
Copyright: © 2025 Li, Wen, Lv, Zhang, Wang and Lang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Zhihong Lang, Biotechnology Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Beijing, China
Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.