Impact of codon optimization on vip3Aa11 gene expression and insecticidal efficacy in maize
文献类型： 外文期刊
作者： Li, Shengyan 1 ; Wen, Ning 1 ; Lv, Wenjie 1 ; Zhang, Mingjun 1 ; Wang, Yinxiao 1 ; Lang, Zhihong 1 ;
作者机构： 1.Biotechnol Res Inst, Chinese Acad Agr Sci, Beijing, Peoples R China
2.Jilin Agr Univ, Coll Plant Protect, Changchun, Peoples R China
3.Chinese Acad Agr Sci, Natl Nanfan Res Inst Sanya, Sanya, Hainan, Peoples R China
关键词：
codon optimization;
期刊名称：FRONTIERS IN PLANT SCIENCE （ 影响因子：4.8； 五年影响因子：5.7 ）
ISSN： 1664-462X
年卷期： 2025 年 16 卷
页码：
收录情况： SCI
摘要： Introduction 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.Methods and results 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.Discussion 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.
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