科学网 - 精选博文 ( ) • 2021-05-17 22:30

研究人员开发出了工程化菌株,以光学控制细菌的运动行为

诸平

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Figure 1. Design principle and application diagram of the engineered strain pactm. (Image by SIAT)  

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Figure 2. Control infection ability of pactm to mice using blue light. (Image by SIAT)  

物理学家组织网phys.org2021517日报道,中国科学院(Chinese Academy of Sciences)深圳先进技术研究所(Shenzhen Institute of Advanced Technology简称SIAT)金帆(Jin Fan音译)领导的研究小组与华中科技大学(Huazhong University of Science and Technology)刘智(Liu Zhi音译)领导的研究小组合作,设计了一种基于铜绿假单胞菌(Pseudomonas aeruginosa)的工程化菌株。此菌株可以光学控制其运动行为以及在宿主上的感染。这项研究的结果于202135日已经在《 ACS合成生物学》(ACS Synthetic Biology)杂志网站发表——Aiguo Xia, Mingjie Qian, Congcong Wang, Yajia Huang, Zhi Liu, Lei Ni, Fan Jin. Optogenetic Modification of Pseudomonas aeruginosa Enables Controllable Twitching Motility and Host InfectionACS Synthetic Biology, 2021, 10(3): 531–541. DOI: 10.1021/acssynbio.0c00559. Publication Date: March 5, 2021.

环磷酸一腺苷(Cyclic adenosine monophosphate简称cAMP)是重要的第二信使(secondary messenger),它控制铜绿假单胞菌的碳代谢,IVa型菌毛(TEP)生物发生(biogenesis)和毒力(virulence)。当cAMP调节细菌中的某种生理功能时,通常会同时调节影响该生理功能的多种成分蛋白的表达水平。

在这项研究中,研究人员通过将光激活腺苷酸环化酶(adenylate cyclase)基因(bPAC)引入细菌,开发出一种具有光依赖性胞内cAMP水平的工程化铜绿假单胞菌(P. aeruginosa)菌株取名为pactm

该工程化菌株在蓝光照射下可可逆地改变其蹭行运动(twitching motility)和宿主感染的能力。在蓝光照射下,pactmcAMP反应启动子的表达量提高了15倍,蹭行运动能力提高了8倍。

裸鼠皮下感染模型显示,pactm感染引起的皮肤损伤面积增加了14倍(见上述图示)。因此,本研究为构建可控感染实验模型提供了一种解决方案。

此外,研究者通过宏观照明模式的设计指导了细菌种群的扩张方向,为微生物间接触依赖相互作用的研究提供了便利。

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Abstract

Cyclic adenosine monophosphate (cAMP) is an important secondary messenger that controls carbon metabolism, type IVa pili biogenesis, and virulence in Pseudomonas aeruginosa. 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 P. aeruginosa strain (termed pactm) with light-dependent intracellular cAMP levels through introducing a photoactivated adenylate cyclase gene (bPAC) into bacteria. On blue light illumination, pactm 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 pactm inoculation was increased 14-fold by blue light, making pactm suitable for applications in controllable bacterial host infection. In addition, we achieved directional twitching motility of pactm colonies through localized light illumination, which will facilitate the studies of contact-dependent interactions between microbial species.




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