Development of a double-antibody sandwich ELISA for quantification of mutated EPSPS gene expression in rice
文献类型： 外文期刊
作者： Luo, Biao 1 ; Zhang, Xianwen 2 ; Wang, Fang 2 ; Wang, Yan 2 ; Wu, Wei 4 ; Lin, Chaoyang 5 ; Rao, Liqun 1 ; Wang, Qiming 1 ;
作者机构： 1.Hunan Agr Univ, Coll Biosci & Biotechnol, Changsha 410128, Hunan, Peoples R China
2.Zhejiang Acad Agr Sci, Inst Virol & Biotechnol, Hangzhou 310021, Zhejiang, Peoples R China
3.Zhejiang Acad Agr Sci, Key Lab Traceabil Agr Genet Modified Organisms, Minist Agr & Rural Affairs, Hangzhou 310021, Zhejiang, Peoples R China
4.Shanghai YouLong Biotech Co Ltd, Shanghai 200063, Peoples R China
5.Zhejiang Univ, Inst Insect Sci, Rural Dev Acad, Hangzhou 310063, Zhejiang, Peoples R China
关键词： 5-Enolpyruvyl-shikimate-3-phosphate synthase (EPSPS); DAS-ELISA; OsmEPSPS detection; Glyphosate-tolerant rice; Glyphosate-tolerant rice
期刊名称：ANALYTICAL BIOCHEMISTRY （ 影响因子：2.5； 五年影响因子：2.7 ）
ISSN： 0003-2697
年卷期： 2025 年 696 卷
页码：
收录情况： SCI
摘要： Glyphosate resistance is a critically important trait for genetically modified (GM) crops. Mutation of the rice EPSPS gene results in a high level of glyphosate resistance, presenting significant potential for the development of glyphosate-tolerant crops. The resistance of rice to glyphosate is correlated with the expression levels of resistance genes. Therefore, developing a convenient, stable, and sensitive method for quantifying the OsmEPSPS protein is crucial for the development of glyphosate-resistant crops. We developed a double-antibody sandwich quantitative ELISA (DAS-ELISA) using a specific monoclonal antibody (mAb) for OsmEPSPS capture and an HRPconjugated anti-OsmEPSPS rabbit polyclonal antibody (pAb). The method could be used to detect OsmEPSPS within a linear range of 16-256 ng/mL with robust intra- and inter-batch duplicability (%CV values: 0.17 %- 7.24 %). OsmEPSPS expression in the transgenic rice lines (54.44-445.80 mu g/g) was quantified using the DASELISA. Furthermore, the expression of the OsmEPSPS gene was validated through Western blotting. This study demonstrated the reliability and stability of the DAS-ELISA for OsmEPSPS detection in GM rice.
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