Tschirhart Tanya, Shukla Vrinda, Kelly Erin E, Schultzhaus Zachary, NewRingeisen Erin, Erickson Jeffrey S, Wang Zheng, Garcia Whitney, Curl Emaleigh, Egbert Robert G, Yeung Enoch, Vora Gary J
American Society for Engineering Education, Postdoctoral Fellowship Program , US Naval Research Laboratory , Washington , DC 20375 , United States of America.
Center for Bio/Molecular Science and Engineering , US Naval Research Laboratory , Washington , DC 20375 , United States of America.
ACS Synth Biol. 2019 Sep 20;8(9):2069-2079. doi: 10.1021/acssynbio.9b00176. Epub 2019 Sep 9.
The fast-growing nonmodel marine bacterium has recently garnered attention as a host for molecular biology and biotechnology applications. In order to further its capabilities as a synthetic biology chassis, we have characterized a wide range of genetic parts and tools for use in . These parts include many commonly used resistance markers, promoters, ribosomal binding sites, reporters, terminators, degradation tags, origin of replication sequences, and plasmid backbones. We have characterized the behavior of these parts in different combinations and have compared their functionality in and . Plasmid stability over time, plasmid copy numbers, and production load on the cells were also evaluated. Additionally, we tested constructs for chemical and optogenetic induction and characterized basic engineered circuit behavior in . The results indicate that, while most parts and constructs work similarly in the two organisms, some deviate significantly. Overall, these results will serve as a primer for anyone interested in engineering and will aid in developing more robust synthetic biology principles and approaches for this nonmodel chassis.
这种快速生长的非模式海洋细菌最近作为分子生物学和生物技术应用的宿主受到了关注。为了进一步提升其作为合成生物学底盘的能力，我们对一系列用于该细菌的遗传元件和工具进行了表征。这些元件包括许多常用的抗性标记、启动子、核糖体结合位点、报告基因、终止子、降解标签、复制起始序列和质粒骨架。我们已经表征了这些元件在不同组合中的行为，并比较了它们在该细菌和其他细菌中的功能。还评估了质粒随时间的稳定性、质粒拷贝数以及细胞上的生产负荷。此外，我们测试了用于化学诱导和光遗传学诱导的构建体，并表征了该细菌中基本工程电路的行为。结果表明，虽然大多数元件和构建体在这两种生物体中的工作方式相似，但有些则有显著差异。总体而言，这些结果将为任何对该细菌工程感兴趣的人提供入门指导，并有助于为这种非模式底盘开发更强大的合成生物学原理和方法。
