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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.
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相关实验视频

Updated: Jun 23, 2025

In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression
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高度敏感和严格的胆诱导基因表达控制器的定向进化.

Yuki Yanai1, Takayuki Hoshino2, Yuki Kimura1

  • 1Department of Applied Chemistry, Faculty of Science and Engineering, Waseda University.

The Journal of general and applied microbiology
|June 16, 2024
PubMed
概括

这项研究增强了用于微生物生产的胆受控基因诱导系统. 改进后的系统实现了严格的基因表达控制,诱导成本显著降低,非常适合工业生物工艺.

关键词:
基因电路是基因电路.感应系统的感应系统.积极的循环循环是积极的.转录因子的转录因子

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科学领域:

  • 合成生物学 合成生物学
  • 代谢工程是代谢工程.
  • 生物技术是生物技术.

背景情况:

  • 基因表达控制器对于微生物生产蛋白质和化学物质至关重要.
  • 现有的系统往往缺乏低成本,安全性和对工业生物工艺的严格控制.
  • 一个先前开发的胆诱导系统需要高诱导度 (~100mM).

研究的目的:

  • 显著提高胆诱导基因表达系统的敏感性.
  • 为了降低用于工业应用的诱导分子的成本.
  • 为了实现更紧密,更有效的基因表达控制.

主要方法:

  • 使用胆进口蛋白 BetT.设计了一个正反循环 (P-loop).
  • 调整了BetT的翻译级别,以优化P循环的强度.
  • 删除了内源性betl基因,以最大限度地减少噪音和提高严格性.

主要成果:

  • 实现了高度活跃和严格的基因表达控制,胆含量减少了100倍.
  • 显示了可诱导胆系统报告的最低基底表达水平.
  • 呈现了最低的激活值和最高的诱导振幅,与PT5等强大的促进剂相当.

结论:

  • 增强的胆诱导系统为基因表达提供了具有成本效益和精确的控制.
  • 该系统克服了以前的局限性,使其能够在大型工业生物工艺中得到更广泛的应用.
  • 微不足道的诱导成本使该系统成为工业生物技术的可行解决方案.