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在转基因Arabidopsi中对外源性和内源性酒精脱酶基因的表达评估
Haoqiang Yu1, Hong Lv1, Fengzhong Lu1
1Maize Research Institute, Sichuan Agricultural University, Chengdu, China.
Frontiers in plant science
|February 6, 2025
概括
来自遥远物种的外源基因,如玉米和大肠杆菌酒精脱酶 (ADH),在阿拉比多普西斯中表现出比内源ADH基因更高的表达,蛋白质积累和酶活性,这表明它们的作物改善潜力更大.
科学领域:
- 植物生物技术 植物生物技术
- 分子生物学分子生物学
- 遗传学 遗传学 是一个
背景情况:
- 作物特征改进通常涉及基因转换,大多数成功的商业活动使用来自遥远物种的外源基因.
- 用于特征增强的内源基因转换在很大程度上仍处于实验阶段.
研究的目的:
- 为了比较转变异源基因与作物内源基因的表达,蛋白质积累和酶活性.
- 为了评估酒精脱酶 (ADH) 基因的有效性从阿拉比多普西斯 (内源),玉米 (外源) 和大肠杆菌 (外源) 在转基因阿拉比多普西斯.
主要方法:
- 克隆的酒精脱酶 (ADH) 编码序列 (CDS) 来自阿拉比多普西斯,玉米和大肠杆菌.
- 构建的表达载体和转换的野生类型的Arabidopsis.
- 使用抗生素查,PCR和DNA再测序查的同卵性T3线.
- 通过RT-qPCR分析基因表达,通过西式涂抹和ELISA进行蛋白质积累,并通过酶试验分析酶活性.
主要成果:
- 转变的外源玉米ADH (ZmADH) 和大肠杆菌ADH (EcADH) 基因表达水平明显高于转变的内源阿拉比多普西斯ADH (AtADH) 基因.
- 蛋白质积累和ADH酶活性在与ZmADH和EcADH转换的阿拉比多普西斯菌株中明显大于与AtADH转换和野生类型对照相比.
- 外源基因转换在增强ADH相关特征方面表现出卓越的表现.
结论:
- 外源基因在转基因作物改良方面比内源基因更有效.
- 玉米和大肠杆菌ADH基因的异质表达为增强作物特征提供了一个有希望的策略.
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