细胞自由蛋白质表达通过重建的转录-翻译系统,由糖代谢提供能量
Gaku Sato1, Shintaro Miyazawa1, Nobuhide Doi1
1Department of Biosciences & Informatics, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522, Japan.
Molecules (Basel, Switzerland)
|July 13, 2024
概括
研究人员重建了糖溶解,并将其与体外转录-翻译 (TX-TL) 相结合. 这种综合系统合成蛋白质和一些自身的酶,为从纯化的成分中构建细胞铺平了道路.
科学领域:
- 合成生物学 合成生物学
- 生物化学 生物化学
- 没有细胞的系统
背景情况:
- 细胞平衡依赖于协调的代谢 (例如,糖解) 和代谢 (例如,转录-翻译,TX-TL).
- 使用纯化组件在体外重建这些基本过程之间的合作是一个重大挑战.
- 了解这种合作是阐明活细胞的设计原理的关键.
研究的目的:
- 通过使用纯化因子,在体外实现糖解的复制及其与转录-翻译 (TX-TL) 的整合.
- 优化糖解和TX-TL的合作功能条件.
- 探索从基本生物组件中创建自我复制系统的潜力.
主要方法:
- 使用糖的糖溶解的复制.
- 复制糖解与PURE系统 (体外TX-TL套件) 的整合.
- 优化关键参数,包括葡萄糖酶和酸盐度.
- 分析ATP消耗效应和使用糖溶性中间体的缓解策略.
主要成果:
- 葡萄糖分解与体外TX-TL成功集成,使蛋白质合成成为可能.
- 在优化条件下,与原始PURE系统相比,达到高达33%的蛋白质合成产量.
- 证明通过糖溶解消耗ATP会抑制TX-TL,可以通过添加糖溶性中间体来减轻.
- 在集成系统中展示了糖解酶的同时合成,这是迈向自我复制的一步.
结论:
- 该研究成功地与体外TX-TL重组并整合了糖解,证明了合作功能.
- 开发的系统代表了从纯化的成分重建活细胞的基本步骤.
- 这个集成系统为研究基本细胞过程和设计最小细胞提供了一个平台.
相关概念视频
01:58Prokaryotic Transcriptional Activators and Repressors
The organization of prokaryotic genes in their genome is notably different from that of eukaryotes. Prokaryotic genes are organized, such that the genes for proteins involved in the same biochemical process or function are located together in groups. This group of genes, along with their regulatory elements, are collectively known as an operon. The functional genes in an operon are transcribed together to give a single strand of mRNA known as polycistronic mRNA.
Transcription of prokaryotic...
Transcription of prokaryotic...
02:11Reporter Genes
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Commonly used reporter...
02:24Ribosome Profiling
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...