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Temperature-sensitive gene modules for synthetic biology applications

用于合成生物学应用的温度敏感基因模块

基本信息

批准号：
41841-2011
负责人：
Nano, Francis
金额：
$ 2.19万
依托单位：
University of Victoria
依托单位国家：
加拿大
项目类别：
Discovery Grants Program - Individual
财政年份：
2011
资助国家：
加拿大
起止时间：
2011-01-01 至 2012-12-31
项目状态：
已结题

来源：
https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=474908
关键词：
Temperature sensitive gene modules synthetic

项目摘要

The proposed research builds on our recent successes of substituting Arctic essential genes into mesophilic pathogenic bacteria to make the latter stably temperature-sensitive (Duplantis, et al., PNAS, 2010). This work was widely recognized as ground-breaking, and we are the leading group in the world carrying out this area of research. For this proposal we outline a research program in which we will discover or develop a broad family of gene cassettes that render microbes temperature-sensitive, and our collection of gene cassettes will allow us to apply our technology to solve many problems in biotechnology and in human and animal health. Two basic approaches will be used. Our current approach relies on bioprospecting to discover essential genes encoded in the genomes of psychrophilic (cold-loving) bacteria. We will continue to use this approach to significantly expand our collection of temperature-sensitive essential genes. We will also test if essential genes from thermophiles (warm-loving) can set the lower temperature growth limit of a mesophile (medium temperature-loving). In the second approach we will develop temperature-sensitive gene cassettes using a variety of synthetic biology approaches. These cassettes will be used to induce the expression of lethal genes or the elimination of an antidote to a toxin that kills a microbe. We expect the family of temperature-sensitive essential genes or cassettes to be versatile enough to be used in any bacterium and some of the cassettes will be useful in the engineering of eukaryotic microbes.

所提出的研究建立在我们最近成功地将北极必需基因替换到嗜温病原菌中以使后者稳定地对温度敏感的基础上（Duplantis等人，PNAS，2010）。这项工作被广泛认为是开创性的，我们是世界上开展这一研究领域的领导小组。对于这项提案，我们概述了一项研究计划，在该计划中，我们将发现或开发一个广泛的基因盒家族，使微生物对温度敏感，我们收集的基因盒将使我们能够应用我们的技术来解决生物技术和人类和动物健康中的许多问题。将采用两种基本方法。我们目前的方法依赖于生物勘探，以发现嗜冷细菌基因组中编码的必需基因。我们将继续使用这种方法来显著扩大我们对温度敏感的必需基因的收集。我们还将测试嗜热菌（嗜暖）的必需基因是否可以设定嗜温菌（嗜中温）的低温生长极限。在第二种方法中，我们将使用各种合成生物学方法开发温度敏感的基因盒。这些盒将用于诱导致命基因的表达或消除杀死微生物的毒素的解毒剂。我们期望温度敏感必需基因或盒的家族足够通用以用于任何细菌，并且一些盒将用于真核微生物的工程化。