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[在大肠杆菌中化学诱导染色体进化的建立和应用Nissle 1917]

Yumei Liu¹, Qingli Zhang², Lijun Shao¹

¹School of Public Health, Shandong Second Medical University, Weifang 261053, Shandong, China.

Sheng wu gong cheng xue bao = Chinese journal of biotechnology

|December 26, 2024

概括

工程细菌 Escherichia coli Nissle 1917 (EcN) 稳定地合成了用于癌症治疗的 5-氨基诺列维氨酸 (5-ALA). 这种方法通过将5-ALA直接输送到瘤细胞来提高光动力学治疗的疗效.

科学领域:

微生物学微生物学
生物技术是生物技术.
癌症治疗癌症治疗

背景情况:

* Escherichia coli Nissle 1917 (EcN) 是一种益生菌菌株,由于其生物相容性和遗传处理性,适合用于细菌癌症治疗.
*目前的EcN工程方法通常依赖于等离子体,这可能导致遗传不稳定,需要选择抗生素.
* 开发稳定的非等离子体表达系统对于可靠的细菌疗法应用至关重要.

研究的目的:

* 设计EcN以稳定合成5 - 氨基联酸 (5-ALA),这是一种用于光动力学治疗的光敏剂.
*通过将治疗基因整合到细菌基因组中来克服基于等离子体的系统的局限性.
*通过化学诱导的染色体进化 (CIChE) 系统来增强5-ALA的产生.

主要方法:

*使用菌体集成将关键的5-ALA合成基因 (hemA^M和hemL) 集成到EcN基因组中.
* 应用化学诱导的染色体进化 (CIChE) 来放大基因拷贝数量并促进5-ALA合成.
*对工程菌株向瘤细胞输送5-ALA的能力进行体外评估,并评估其治疗效果.

主要成果:

*成功将5-ALA合成基因集成到EcN基因组中.
* CIChE显著增加了hemA^M和hemL的副本数量,导致稳定的5-ALA生产.
*在体外实验证实,工程 EcN 菌株有效地将5-ALA 传递给瘤细胞,抑制其生长.

结论:

* 建立了一个稳定的,基因组集成的系统,用于在EcN中合成5-ALA,克服了等离子体不稳定性问题.
* 经过工程设计的EcN菌株证明了其作为光动力学癌症治疗的治疗剂的潜力.
*这项研究为开发使用转基因益生菌菌株的先进细菌疗法提供了基础.

关键词:

5 - 氨基氨基氨基氨基酸.大肠杆菌 Escherichia coli 尼斯尔 1917 年化学诱导的染色体进化 (CIChE)菌体集成技术的方法

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