Single live-bacterial cell assay of promoter activity and regulation

Jun Teramoto, Yoko Yamanishi, El Shimy H. Magdy, Akiko Hasegawa, Ayako Kori, Masahiro Nakajima, Fumihito Arai, Toshio Fukuda, Akira Ishihama

研究成果: ジャーナルへの寄稿記事

10 引用 (Scopus)

抄録

Laboratory cultures of a single species of bacteria harboring the same genetic background include heterogeneous cell populations, each differing in apparent morphology and physiology, as found in natural environments. To get insights into difference in the genome expression between individual cells, we constructed various types of the cell chip for monitoring the growth and fate of individual bacterial cells. Immobilization of portions of Escherichia coli culture within these cell chips was established after raising the local temperature in the presence of poly-(N-isopropylacrylamide) (PNIPAAm). The newly developed cell-chip system allows the investigation of activity and regulation of green fluorescent protein (GFP)-fused promoter in single live-bacterial cells for prolonged time under controlled culture conditions. Using this single-cell observation system, we succeeded, for the first time, the real-time single-cell assay of promoter activity of the E. coli gcl gene encoding glyoxylate carboligase as a model system, and the kinetics of gcl induction by an effector glyoxylate. Marked heterogeneity was found in the expression level of the gcl promoter. The heterogeneity in gcl promoter activity was, however, confirmed by Flow cytometry of suspension cultures. Our success provides an experimental system for the increased demand of single-cell biology in bacterial studies.

元の言語英語
ページ(範囲)1111-1122
ページ数12
ジャーナルGenes to Cells
15
発行部数11
DOI
出版物ステータス出版済み - 11 1 2010

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tartronate-semialdehyde synthase
Escherichia coli
Green Fluorescent Proteins
Immobilization
Cell Biology
Suspensions
Flow Cytometry
Cell Culture Techniques
Observation
Genome
Bacteria
Temperature
Growth
Population
Genes
Genetic Background
glyoxylic acid
poly-N-isopropylacrylamide

All Science Journal Classification (ASJC) codes

  • Genetics
  • Cell Biology

これを引用

Teramoto, J., Yamanishi, Y., Magdy, E. S. H., Hasegawa, A., Kori, A., Nakajima, M., ... Ishihama, A. (2010). Single live-bacterial cell assay of promoter activity and regulation. Genes to Cells, 15(11), 1111-1122. https://doi.org/10.1111/j.1365-2443.2010.01449.x

Single live-bacterial cell assay of promoter activity and regulation. / Teramoto, Jun; Yamanishi, Yoko; Magdy, El Shimy H.; Hasegawa, Akiko; Kori, Ayako; Nakajima, Masahiro; Arai, Fumihito; Fukuda, Toshio; Ishihama, Akira.

:: Genes to Cells, 巻 15, 番号 11, 01.11.2010, p. 1111-1122.

研究成果: ジャーナルへの寄稿記事

Teramoto, J, Yamanishi, Y, Magdy, ESH, Hasegawa, A, Kori, A, Nakajima, M, Arai, F, Fukuda, T & Ishihama, A 2010, 'Single live-bacterial cell assay of promoter activity and regulation', Genes to Cells, 巻. 15, 番号 11, pp. 1111-1122. https://doi.org/10.1111/j.1365-2443.2010.01449.x
Teramoto J, Yamanishi Y, Magdy ESH, Hasegawa A, Kori A, Nakajima M その他. Single live-bacterial cell assay of promoter activity and regulation. Genes to Cells. 2010 11 1;15(11):1111-1122. https://doi.org/10.1111/j.1365-2443.2010.01449.x
Teramoto, Jun ; Yamanishi, Yoko ; Magdy, El Shimy H. ; Hasegawa, Akiko ; Kori, Ayako ; Nakajima, Masahiro ; Arai, Fumihito ; Fukuda, Toshio ; Ishihama, Akira. / Single live-bacterial cell assay of promoter activity and regulation. :: Genes to Cells. 2010 ; 巻 15, 番号 11. pp. 1111-1122.
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