Spatiotemporal mapping of gene expression to reveal mechanisms of cellular differentiation in E. coli biofilms

基因表达的时空图谱揭示大肠杆菌生物膜细胞分化机制

• 批准号: 21K05341
• 负责人: Robert Martin
• 金额: $ 2.58万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2021
• 资助国家: 日本
• 起止时间: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-21K05341/
• 关键词: biofilm; E. coli; imaging system; gene expression; proteome; cell motility; surface properties; nutrients; differentiation; Escherichia coli

During the second fiscal year we have progressed on multiple fronts.1) We started and progressed with spatial analysis of protein expression by sampling biofilms at different radial locations followed by ongoing proteome analysis.2) We characterized further expression of selected promoters activity under different conditions. To normalize the fluorescence results we have been testing a secondary fluorescent protein whose expression is constitutive.3) We have been characterizing the effect of glucose addition on biofilm development and our results suggest that an unsuspected metabolic imbalance may be important in the process.4) We have developed imaging systems based on Raspberry Pi computers and cameras to collect time-lapse images in dual fluorescence mode in addition to bright field.

在第二个财政年度，我们在多个方面取得了进展。1)我们开始并进行了蛋白质表达的空间分析，通过在不同的径向位置取样生物膜，然后进行蛋白质组学分析。2)我们进一步表征了所选启动子活性在不同条件下的表达。为了使荧光结果正常化，我们一直在测试一种二级荧光蛋白，其表达是组成的。3)我们一直在描述葡萄糖添加对生物膜发育的影响，我们的结果表明，在这个过程中，一种意想不到的代谢失衡可能是重要的。4)我们开发了基于树莓派电脑和相机的成像系统，除了亮场外，还可以采集双荧光模式的延时图像。

项目成果

Exploring developmental processes in growing E. coli biofilms

探索大肠杆菌生物膜的发育过程

• 发表时间: 2021
• 作者: Fujino Y. Sakai;R. and Robert M.;Robert M.;Yukiko Fujino
• 通讯作者: Yukiko Fujino

Opening and closing the box of bacterial metabolic function: from enzyme discovery to adaptive evolution and biofilms using E. coli.

打开和关闭细菌代谢功能的盒子：从酶的发现到使用大肠杆菌的适应性进化和生物膜。

• 发表时间: 2022
• 作者: Maruyama T.;Sumi S.;Kobayashi M.;Ebuchi T.;Kanesaki Y.;Yoshikawa H.;Ueda K.;and Takano H.;Robert M.
• 通讯作者: Robert M.

Exploring transitions in metabolic state during the development of bacterial biofilms.

探索细菌生物膜发育过程中代谢状态的转变。

• 发表时间: 2023
• 作者: Fujino Y. Sakai;R. and Robert M.
• 通讯作者: R. and Robert M.