CAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, PR China.
Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, PR China.
ACS Synth Biol. 2021 Mar 19;10(3):531-541. doi: 10.1021/acssynbio.0c00559. Epub 2021 Mar 5.
Cyclic adenosine monophosphate (cAMP) is an important secondary messenger that controls carbon metabolism, type IVa pili biogenesis, and virulence in . Precise manipulation of bacterial intracellular cAMP levels may enable tunable control of twitching motility or virulence, and optogenetic tools are attractive because they afford excellent spatiotemporal resolution and are easy to operate. Here, we developed an engineered strain (termed ) with light-dependent intracellular cAMP levels through introducing a photoactivated adenylate cyclase gene () into bacteria. On blue light illumination, displayed a 15-fold increase in the expression of the cAMP responsive promoter and an 8-fold increase in its twitching activity. The skin lesion area of nude mouse in a subcutaneous infection model after 2-day inoculation was increased 14-fold by blue light, making suitable for applications in controllable bacterial host infection. In addition, we achieved directional twitching motility of colonies through localized light illumination, which will facilitate the studies of contact-dependent interactions between microbial species.
环磷酸腺苷 (cAMP) 是一种重要的次级信使，它控制着碳代谢、IVa 型菌毛生物发生和毒力。精确控制细菌细胞内 cAMP 水平可能实现对蠕动运动或毒力的可调控制，而光遗传学工具很有吸引力，因为它们具有出色的时空分辨率，并且易于操作。在这里，我们通过将光激活的腺苷酸环化酶基因 () 引入细菌，开发了一种具有光依赖性细胞内 cAMP 水平的工程菌 (命名为 )。在蓝光照射下， 的 cAMP 反应启动子表达增加了 15 倍，其蠕动活性增加了 8 倍。在皮下感染模型中，经过 2 天的 接种后，裸鼠的皮肤损伤面积增加了 14 倍，这使得 适合用于可控细菌宿主感染的应用。此外，我们通过局部光照实现了 菌落的定向蠕动运动，这将有助于研究微生物种间的接触依赖性相互作用。
