Temperature-sensitive gene modules for synthetic biology applications

点击翻译按钮获取中文摘要

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.