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在转基因作物中快速检测复合大米EPSPS的免疫染色带的开发

Biao Luo¹, Fang Wang², Qiming Wang³

¹College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha, 410128, China; Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021, China.

Analytical biochemistry

|June 8, 2025

概括

一种新的免疫染色条纹 (ICS) 测试可以快速检测OsmEPSPS蛋白,这对于转基因玉米和大豆的草甘耐药性至关重要. 这种敏感和特定的测试有助于识别耐药植物和分析EPSPS基因同质性.

科学领域:

农业生物技术农业生物技术
分子生物学分子生物学
生物化学生物化学

背景情况:

耐草甘作物利用EPSPS基因进行除草剂耐受性.
这种TIPS-OsEPSPS基因赋予了大米的草甘耐药性.
开发具有增强草甘耐药性的玉米和大豆等作物是农业的一个关键目标.

研究的目的:

使用OsmEPSPS基因开发抗草甘的玉米和大豆.
为特定的OsmEPSPS蛋白质检测设计一种快速免疫染色带 (ICS) 测试.
评估开发的ICS测试的灵敏度,特异性和稳定性.

主要方法:

利用OsmEPSPS基因设计抗草甘玉米和大豆.
开发了一种使用双抗体三明治格式的免疫染色体条纹 (ICS) 测试.
使用单克隆抗体 (mAb #9和mAb #15) 来捕获和检测OsmEPSPS蛋白质.

主要成果:

ICS测试显示,在10分钟内检测到0.031μg/ml的检测极限.
快速检测 (0.817μg/mL在1分钟内) 在Eleusine indica和OsmEPSPS过度表达的植物中实现.
ICS显示高特异性,灵敏度,稳定性 (R2 = 0.982),并检测出同源的EPSPS蛋白.

结论:

成功开发了一种敏感和特定的ICS测试,用于OsmEPSPS检测.
该ICS有助于半定量分析和快速识别OsmEPSPS过度表达植物.
开发的ICS对于作物识别和EPSPS基因同质性的植物遗传学分析都是有价值的.

关键词:

5 - 乙醇pyruvyl - 石基酸 - 3 - 酸盐合成酶 (EPSPS)对草甘耐药的作物种植.免疫染色图带 (ICS) 是指免疫染色图带.快速检测快速检测快速检测快速检测

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