Document Detail

Single live-bacterial cell assay of promoter activity and regulation.
MedLine Citation:
PMID:  20964794     Owner:  NLM     Status:  MEDLINE    
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.
Jun Teramoto; Yoko Yamanishi; El-Shimy H Magdy; Akiko Hasegawa; Ayako Kori; Masahiro Nakajima; Fumihito Arai; Toshio Fukuda; Akira Ishihama
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-10-21
Journal Detail:
Title:  Genes to cells : devoted to molecular & cellular mechanisms     Volume:  15     ISSN:  1365-2443     ISO Abbreviation:  Genes Cells     Publication Date:  2010 Nov 
Date Detail:
Created Date:  2010-10-27     Completed Date:  2011-01-31     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9607379     Medline TA:  Genes Cells     Country:  England    
Other Details:
Languages:  eng     Pagination:  1111-22     Citation Subset:  IM    
Copyright Information:
© 2010 The Authors. Journal compilation © 2010 by the Molecular Biology Society of Japan/Blackwell Publishing Ltd.
Department of Frontier Bioscience, Hosei University, Koganei, Tokyo, Japan.
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MeSH Terms
Acrylamides / chemistry
Bacteriological Techniques
Biological Assay / methods*
Escherichia coli / genetics*,  growth & development,  metabolism
Escherichia coli Proteins / genetics
Flow Cytometry
Gene Expression Regulation, Bacterial*
Green Fluorescent Proteins / biosynthesis,  genetics,  metabolism
Promoter Regions, Genetic*
Repressor Proteins / genetics
Reg. No./Substance:
0/Acrylamides; 0/Escherichia coli Proteins; 0/GclR protein, E coli; 0/Repressor Proteins; 147336-22-9/Green Fluorescent Proteins; 2210-25-5/N-isopropylacrylamide

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