蛋白质拼接的光激活与光中的快速蛋白质
1Department of Chemistry, University of California, Riverside , 501 Big Springs Road, Riverside, California 92521, United States.
Journal of the American Chemical Society
|February 4, 2015
概括
研究人员设计了一种光激活的整蛋白,用于哺乳动物细胞中的光遗传控制. 这种工具可以精确控制蛋白质拼接和使用光的功能,开辟了生物技术的新途径.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 视觉遗传学 视觉遗传学
背景情况:
- 蛋白质间接拼接是一种强大的工具,具有多样化的生物技术应用.
- 使用外部刺激,如光,控制蛋白质活动对于先进的生物研究至关重要.
研究的目的:
- 设计一种可光激活的蛋白质,用于活哺乳动物细胞中.
- 通过光诱导激活来证明控制蛋白质拼接和功能的能力.
主要方法:
- 在Nostoc点形 (Npu) DnaE中基因引入了一种光的囊蛋白.
- 通过将光中的整蛋白插入mCherry和人类Src氨酸激酶中,制造出非活性的嵌合蛋白蛋白.
- 利用光诱导的光化学反应来重新激活整蛋白,并触发哺乳动物细胞中的蛋白质拼接.
主要成果:
- 成功创建了一种与哺乳动物细胞相容的可光激活蛋白质.
- 证明了因特因和随后的蛋白质拼接的光诱导的活性化.
- 通过光成像和激酶活性测试观察到活性mCherry和Src蛋白的形成.
结论:
- 基因编码的光内蛋白作为一个多功能的光遗传工具.
- 这项技术可以有效地对蛋白质结构和功能进行光控制.
- 能够精确的时间和空间调节活细胞中的生物过程.
相关概念视频
Tagging and Fusion Proteins
9.0K
Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
9.0K
Conservative Site-specific Recombination and Phase Variation
7.4K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
7.4K


