Visualization, sorting, and transcriptome analysis of Escherichia coli subpopulations with different generations of poles.

2015 Fiscal Year Final Research Report

• Project/Area Number: 26660078
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Applied microbiology
• Research Institution: National Institute of Advanced Industrial Science and Technology
• Principal Investigator: SUYAMA Tetsushi 国立研究開発法人産業技術総合研究所, バイオメディカル研究部門, 主任研究員 (10357311)
• Co-Investigator(Kenkyū-buntansha): MIZUNO Takafumi 国立研究開発法人産業技術総合研究所, バイオメディカル研究部門, 主任研究員 (40344163) ; NODA Naohiro 国立研究開発法人産業技術総合研究所, バイオメディカル研究部門, 研究グループリーダー (70415727)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: 大腸菌 / 細胞分裂 / サブポピュレーション / 蛍光蛋白質 / 光変換 / LED / バイオイメージング / 分取

Outline of Final Research Achievements

Rod-shaped bacteria (e.g., Escherichia coli) are the gathering of subpopulations with multi-generations of their older poles and their physiological traits are different to each other. In this study, we constructed an E. coli with a photoconvertible fluorescent protein conjugated to a chemoreceptor protein Tar localized to their poles, and the generations of poles were successfully visualized. We have further investigated the method of sorting, and established a protocol of photo-labeling without damaging the cells. Specific weight of the cells was found to be a promising method for differentiating the subpopulations with different generations of poles. We have also composed an automatic cell-sorting system utilizing a microfluidic chip, a fluorescent microscope, and a photonic multichannel analyzer for collecting more variety of subpopulations to clarify the mechanisms of modulating growth rate and entering VBNC state in the subsequent studies.

Free Research Field

環境微生物学