导入自然和非自然核酸酸盐到细菌中的工具
Aaron W Feldman1, Emil C Fischer1, Michael P Ledbetter1
1Department of Chemistry, The Scripps Research Institute , 10550 North Torrey Pines Road, La Jolla, California 92037, United States.
Journal of the American Chemical Society
|January 18, 2018
概括
研究人员开发了一种新方法,使用专门的输送器将核酸三酸盐导入细胞. 这一突破有助于研究自然过程,
科学领域:
- 分子生物学
- 合成生物学
- 生物化学
背景情况:
- 核三酸盐是重要的细胞分子,但由于严格的监管和标记或修改的困难,它们的研究具有挑战性.
- 合成生物学旨在开发用于特定细胞功能的非天然核三酸盐.
研究的目的:
- 探索核酸三酸转运器PtNTT2在细胞中引入三酸的普遍性和有用性.
- 进行PtNTT2的结构-活性关系研究,以了解其基质特异性.
主要方法:
- 在大肠杆菌中重新组合的Phaeodactylum tricornutum核酸三转运体 (PtNTT2).
- 竞争性吸收抑制试验以特征转运物的活性与修饰核酸三酸盐.
- 液体染色体-双重质谱 (LC-MS/MS) 试验用于直接测量三酸盐进口.
- 放射性标记或2'-改性三酸盐的进口以及它们在DNA和RNA中的含量定量.
主要成果:
- PtNTT2促进了各种天然和改造的核酸三酸盐进入细胞.
- 结构-活性关系研究阐明了载体对核基,糖和三酸盐修饰的偏好.
- 已成功将进口的修饰三酸盐纳入DNA和RNA.
结论:
- PtNTT2提供了将所需的核酸三酸盐引入细胞的一般方法.
- 这种方法对多种分子和合成生物学应用有价值,包括研究核酸代谢和开发新疗法.
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