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Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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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.
The recognition sites for Cre recombinase called LoxP...
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CRISPR

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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CRISPR and crRNAs

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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Homologous Recombination02:31

Homologous Recombination

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Updated: Jul 9, 2025

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
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Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

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基于CRISPR基基编辑器的向随机突变发生 (BE-TRM) 工具箱用于定向进化.

Rahul Mahadev Shelake1, Dibyajyoti Pramanik1, Jae-Yean Kim2

  • 1Division of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju 52828, Korea.

BMB reports
|December 6, 2023
PubMed
概括
此摘要是机器生成的。

使用基编辑-TRM (BE-TRM) 工具的定向进化 (DE) 能够在复杂的基因组中实现高效的遗传变异. 这些先进的工具克服了以前的局限性,促进了多细胞生物中的DE,用于新型蛋白质和代谢途径的工程.

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科学领域:

  • 分子生物学分子生物学
  • 生物技术是生物技术.
  • 遗传学 是一个遗传学.

背景情况:

  • 定向进化 (DE) 对于产生具有功能改进的遗传变异至关重要.
  • 传统的DE方法仅限于更简单的生物,如细菌和酵母.
  • 长生命周期和低突变率等障碍阻碍了多细胞生物中的DE.

研究的目的:

  • 审查最近在DE的基础编辑-TRM (BE-TRM) 工具中取得的进展.
  • 突出BE-TRM在多细胞系统中的扩大范围和效率.
  • 讨论BE-TRM在生物研究中的应用和未来方向.

主要方法:

  • 基于CRISPR/Cas和DNA脱氨酶的工具已经开发出来,以克服DE的局限性.
  • 基数编辑-TRM (BE-TRM) 允许有针对性的基数替换和序列随机化.
  • 创建和选突变库是DE过程的核心.

主要成果:

  • BE-TRM工具显著提高了DE计划的范围和效率.
  • 这些工具在多细胞生物的原生遗传环境中促进DE.
  • BE-TRM支持蛋白质工程和代谢途径优化的持续分子进化.

结论:

  • BE-TRM代表了DE方法学的重大进步.
  • 它为设计新功能和预测抗生素耐药性等特征提供了一个强大的平台.
  • 对BE-TRM工具的进一步改进将继续扩大其生物应用.