The Geobacillus Plasmid Set: A Modular Toolkit for Thermophile Engineering.
The Geobacillus Plasmid Set: A Modular Toolkit for Thermophile Engineering.
复制标题
DOI：
10.1021/acssynbio.5b00298
复制
发表时间：
2016-07
影响因子：
4.7
通讯作者：
Benjamin Reeve;E. Martínez-Klimova;Joachim De Jonghe;D. Leak;T. Ellis
中科院分区：
文献类型：
--
作者：
Benjamin Reeve;E. Martínez-Klimova;Joachim De Jonghe;D. Leak;T. Ellis
热葡糖苷酸土芽孢杆菌是一种具有工业意义的革兰氏阳性嗜热菌，其生长迅速，并可利用多种植物来源的原料。它是一种有吸引力的底盘生物高温生物技术和合成生物学应用，但目前受到缺乏可用的遗传工具的限制。在这里，我们描述了一套模块化穿梭载体，包括启动子库和报告蛋白。紧凑型质粒由可互换的模块组成，用于在大肠杆菌中的分子克隆和在G. thermoglucosidasius和其他土芽孢杆菌属物种。模块包括两个复制起点，两个选择性标记和三个报告基因表达的表征蛋白。为了从这些质粒中微调异源表达，我们包括一个特征启动子库和测试核糖体结合位点设计。总之，这些基因表达工具和标准化质粒组可以促进模块化和部分交换，使土芽孢杆菌成为合成生物学的嗜热底盘。
Geobacillus thermoglucosidasius is a Gram-positive thermophile of industrial interest that exhibits rapid growth and can utilize a variety of plant-derived feedstocks. It is an attractive chassis organism for high temperature biotechnology and synthetic biology applications but is currently limited by a lack of available genetic tools. Here we describe a set of modular shuttle vectors, including a promoter library and reporter proteins. The compact plasmids are composed of interchangeable modules for molecular cloning in Escherichia coli and stable propagation in G. thermoglucosidasius and other Geobacillus species. Modules include two origins of replication, two selectable markers and three reporter proteins for characterization of gene expression. For fine-tuning heterologous expression from these plasmids, we include a characterized promoter library and test ribosome binding site design. Together, these gene expression tools and a standardized plasmid set can facilitate modularity and part exchange to make Geobacillus a thermophile chassis for synthetic biology.