一种快速和简化的方法来隔离基于作物中的生物素-阿维丁结合亲和关系的特定调节剂
Xiaojian Qin1,2, Yuntong Li1, Cuiping Li1
1College of Life Sciences, Chongqing Normal University, Chongqing 401331, China.
Journal of agricultural and food chemistry
|December 20, 2023
概括
我们开发了一种新的基于DNA的生物素和阿维丁系统 (DBAS),以识别调节作物基因的转录因子 (TF). 这种方法成功地确定了PIP3和PIP19作为米OsNRT1.1B的功能调节剂,证明了其在作物研究中的实用性.
科学领域:
- 植物生物学 植物生物学
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
背景情况:
- 转录因子 (TFs) 对于调节作物中的基因表达至关重要.
- 确定控制作物生长和发展的特定TF对于农业进步至关重要.
研究的目的:
- 开发和验证一种新的基于DNA的生物素和阿维丁系统 (DBAS),用于识别作物的上游转录因子.
- 使用开发的DBAS方法选和识别米OsNRT1.1B基因的调节者.
主要方法:
- 使用共沉试验构建和验证生物素和素系统 (BAS).
- 通过体内和体外测试来证明基于DNA的BAS (DBAS) 的可行性.
- 克隆米的OsNRT1.1B促销者和选潜在的监管因素.
- 通过电泳移动转移试验 (EMSA),酵母单杂交和光酶活性试验验候选调节者的验证.
主要成果:
- 新的DBAS方法在作物中成功建立和验证了TF识别.
- 分析了大米OsNRT1.1B促进体,并确定了候选调节体PIP3和PIP19.
- 证实PIP3和PIP19对酸盐有快速反应,而PIP3的过度表达抑制了生长,这表明它们的功能作用.
结论:
- 开发的DBAS方法有效地隔离了调节作物的特定基因的功能转录因子.
- 已识别的调节剂PIP3和PIP19在植物对酸盐的反应和整体生长中起着重要作用.
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