Single Live-Bacterial Cell Assay of Promoter Activity and Regulation

Teramoto, Jun; Hasegawa, Akiko; Shohata, Eri; Kori, Ayako; Magdy, El Shimy H.; Nakajima, Masahiro; Lin, Yu‐Ching; Yamanishi, Yoko; Magariyama, Yukio; Arai, Fumihito; Fukuda, Toshio; Ishihama, Akira

doi:10.1109/mhs.2008.4752467

Cited by 4 publications

(3 citation statements)

References 38 publications

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“…Only a clear overexpression of the glycerate biosynthesis gcl operon was observed in WDL7 during consortium growth. This operon was shown to be induced by glyoxylate in Escherichia coli (Teramoto et al., ), suggesting that WDL7 senses a higher concentration of glyoxylate when grown in consortium conditions and thus that glyoxylate is a candidate metabolite that WDL7 could receive during consortium growth. As WDL7 does not show a strong change in regulation of metabolic pathways between growth conditions, 3,4‐DCA seems to remain the main carbon and energy source of WDL7 both in monoculture and consortium biofilm conditions.…”

Section: Discussionmentioning

confidence: 99%

Molecular processes underlying synergistic linuron mineralization in a triple‐species bacterial consortium biofilm revealed by differential transcriptomics

et al. 2018

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The proteobacteria Variovorax sp. WDL1, Comamonas testosteroni WDL7, and Hyphomicrobium sulfonivorans WDL6 compose a triple‐species consortium that synergistically degrades and grows on the phenylurea herbicide linuron. To acquire a better insight into the interactions between the consortium members and the underlying molecular mechanisms, we compared the transcriptomes of the key biodegrading strains WDL7 and WDL1 grown as biofilms in either isolation or consortium conditions by differential RNAseq analysis. Differentially expressed pathways and cellular systems were inferred using the network‐based algorithm PheNetic. Coculturing affected mainly metabolism in WDL1. Significantly enhanced expression of hylA encoding linuron hydrolase was observed. Moreover, differential expression of several pathways involved in carbohydrate, amino acid, nitrogen, and sulfur metabolism was observed indicating that WDL1 gains carbon and energy from linuron indirectly by consuming excretion products from WDL7 and/or WDL6. Moreover, in consortium conditions, WDL1 showed a pronounced stress response and overexpression of cell to cell interaction systems such as quorum sensing, contact‐dependent inhibition, and Type VI secretion. Since the latter two systems can mediate interference competition, it prompts the question if synergistic linuron degradation is the result of true adaptive cooperation or rather a facultative interaction between bacteria that coincidentally occupy complementary metabolic niches.

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Section: Discussionmentioning

confidence: 99%

Molecular processes underlying synergistic linuron mineralization in a triple‐species bacterial consortium biofilm revealed by differential transcriptomics

et al. 2018

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“…Micromanipulation has been applied to single bacterial cells since 1960s (Nossal et al, 1964;Wood, 1967). In mechanical micromanipulation, single cells are individually captured from a population and transferred using a micropipette (Anis et al, 2010(Anis et al, , 2011Ashida et al, 2010;Gao et al, 2011;Roeder et al, 2010;Shi et al, 2011;Teramoto et al, 2010;Tsang et al, 2006). The isolated single cells can be subsequently used for different applications such as cultivation or gene expression analysis.…”

Section: Micromanipulationmentioning

confidence: 99%

Measuring gene expression in single bacterial cells: recent advances in methods and micro-devices

Shi

Gao

Wang

et al. 2014

Critical Reviews in Biotechnology

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Populations of bacterial cells that grow under the same conditions and/or environments are often considered to be uniform and thus can be described by ensemble average values of their physiologic, phenotypic, genotypic or other parameters. However, recent evidence suggests that cell-to-cell differences at the gene expression level could be an order of magnitude greater than previously thought even for isogenic bacterial populations. Such gene expression or transcriptional-level heterogeneity determines not only the fate of individual bacterial cells in a population but could also affect the ultimate fate of the population itself. Although techniques for single-cell gene expression measurement in eukaryotic cells have been successfully implemented for a decade or so, they have only recently become available for single bacterial cells. This is due to the difficulty of efficient lysis of most bacterial cells, as well as short half-life time (low stability) of bacterial mRNA. In this article, we review the recent progress and challenges associated with analyzing gene expression levels in single bacterial cells using various semi-quantitative and quantitative methods. In addition, a review of the recent progress in applying microfluidic devices to isolate single bacterial cells for gene expression analysis is also included.

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“…Manipulation and processing technology of micro organisms and cells have been one of the important subjects to be developed with the development of the micro-nano processing technology and gene technology and neurology recently [7][8][9][10]. However they require a special environment such as in the medium (water) and under atmospheric pressure and which limited the device to be operated.…”

Section: Introductionmentioning

confidence: 99%

Local ablation by micro-electric knife

Yamanishi

Kuriki

Sakuma

et al. 2012

2012 7th IEEE International Conference on Nano/Micro Engineered and Molecular Systems (NEMS)

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We have firstly succeeded in enucleation of oocyte by using micro-electric knife. The discharged output power and conductive area were controlled to adapt the cellular-order ablation under water and under atmospheric pressure environment. The local area ablation was obtained by the glass shell insulation around the copper wire with silver paste, and which prevent the bubble generation when it was discharged under the water. This fabricated sharp electric knife has a designed small space at the end of the knife between the wire and the glass tip, and which contribute to stabilize the electric discharge. The width of the ablation region was successfully reduced to more than a half compare to the conventional wire electrode, by using the robust glass shell insulation and a single micro-scale bubble injected from inside of the electrode. Moreover, the nucleus dyed by Hoechst was also removed successfully with limited damage region which is the difficult task for manual operation with glass capillary. This low cost micro electric knife can fabricate any objective material under various environments and it has a possibility to contribute to a new fabrication method in micro-nano bioengineering field.

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Single Live-Bacterial Cell Assay of Promoter Activity and Regulation

Cited by 4 publications

References 38 publications

Molecular processes underlying synergistic linuron mineralization in a triple‐species bacterial consortium biofilm revealed by differential transcriptomics

Molecular processes underlying synergistic linuron mineralization in a triple‐species bacterial consortium biofilm revealed by differential transcriptomics

Measuring gene expression in single bacterial cells: recent advances in methods and micro-devices

Local ablation by micro-electric knife

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