在合成烟草的 хлоропласт操作子中,在广泛的动态范围内对基因表达进行后转录调整
Qiguo Yu1, Tarinee Tungsuchat-Huang1, Alexander Ioannou1
1Waksman Institute of Microbiology, Rutgers University, Piscataway, New Jersey, 08854, USA.
The Plant journal : for cell and molecular biology
|July 20, 2024
概括
这项研究展示了如何使用调节元件精确地控制叶绿体中的基因表达. 研究人员在广泛的范围内调整了记者基因输出,增强了合成生物学应用.
科学领域:
- 植物分子生物学 植物分子生物学
- 合成生物学 合成生物学
- 基因工程是一种基因工程.
背景情况:
- 合成操作子中最佳平衡的基因表达需要多功能调节元素.
- 塑的atpH 5'非翻译区域 (UTR) 含有一种对PPR10的结合位,PPR10是一种后转录激活剂.
研究的目的:
- 用5'UTR.atpH的变体来研究烟草的叶绿体中gfp记者基因的表达.
- 探索PPR10和UTR变异如何影响记者基因表达水平.
- 扩大塑合成生物学工具包,需要多基因表达.
主要方法:
- 在烟草的叶绿体中利用了gfp记者基因表达.
- 采用塑性质atpH 5' UTR的变体,具有不同的设计背景.
- 在UTR上游引入了转移RNA (tRNA),以评估其对基因表达的影响.
主要成果:
- 内源性烟草PPR10与烟草和玉米的atpH 5' UTRs赋予了不同的报告者激活水平.
- 高GFP表达与单基斯特基GFP转录的稳定不相关.
- 在UTR上游加入一个tRNA显著降低了gfp mRNA和蛋白质水平,可能是由于RNA的快速降解.
- 一种tRNA和突变的atpH 5' UTR的组合导致了异常低的转基因表达.
- 实现了可调节的记者基因表达,在总可溶性细胞蛋白的0.02%至25%之间.
结论:
- PPR10激活翻译独立于转录稳定.
- 这项研究提供了一种方法来调整跨广谱的记者基因表达.
- 研究结果支持在塑合成生物学中使用来自异质物种的cis元素.
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