Exploring the Potential of Networked Directed Evolution Based on Novel LacI/effector Pairs

探索基于新型 LacI/效应器对的网络化定向进化的潜力

• 批准号: BB/J008214/1
• 负责人: Florian Hollfelder
• 金额: $ 41.32万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2012
• 资助国家: 英国
• 起止时间: 2012 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FJ008214%2F1
• 关键词: Exploring; Potential; Networked; Directed; Evolution

Understanding of the evolution of functional proteins remains a daunting challenge, despite intense research efforts in basic and applied research. Nature's ability to create functional proteins is still unrivalled, raising the question in which respect we can can improve biomimetic efforts in directed evolution to make laboratory evolution more powerful. One key difference is the context in which evolution occurs: laboratory evolution usually deals with enzymes as single species, but natural enzymes are typically embedded in regulated networks and interdependent pathways. This project attempts to build simple networks in which expression of a coupled reporter marks E. coli cells that successfully express a new, functional protein and renders them selectable. This approach could expand the reaction classes that are accessible for directed evolutino, as current laboratory evolution is primarily targeted at functions that are assayed directly, e.g. by generating a fluorescent product. We hope to develop a method that will make directed evolution more versatile and efficient, but also provide insight into how network regulation might have contributed to natural evolution.

尽管在基础研究和应用研究中进行了激烈的研究工作，但对功能蛋白的演变的理解仍然是一个艰巨的挑战。大自然创建功能蛋白的能力仍然是无与伦比的，这提出了一个问题，我们可以在指导进化中改善仿生努力，以使实验室进化更强大。一个关键区别是发生进化的背景：实验室进化通常将酶作为单个物种涉及酶，但是天然酶通常嵌入受调节的网络和相互依存的途径中。该项目试图构建简单的网络，在该网络中，耦合记者的表达标记了大肠杆菌细胞，这些大肠杆菌细胞成功地表达了一种新的功能蛋白质并使其可选。这种方法可以扩大针对定向Evolutino的反应类别，因为当前的实验室进化主要针对直接测定的功能，例如通过生成荧光产品。我们希望开发一种将使定向进化更加用途和高效的方法，同时也可以深入了解网络调节如何促进自然进化。

项目成果

A single mutation in the core domain of the lac repressor reduces leakiness.

• DOI: 10.1186/1475-2859-12-67
• 发表时间: 2013-07-08
• 期刊: Microbial cell factories
• 影响因子: 6.4
• 作者: Gatti-Lafranconi P;Dijkman WP;Devenish SR;Hollfelder F
• 通讯作者: Hollfelder F