Single-cell microfluidics to study the effects of genome deletion on bacterial growth behavior

Yuan, X. , Couto, J. M., Glidle, A., Song, Y. , Sloan, W. and Yin, H. (2017) Single-cell microfluidics to study the effects of genome deletion on bacterial growth behavior. ACS Synthetic Biology, 6(12), pp. 2219-2227. (doi: 10.1021/acssynbio.7b00177) (PMID:28844132)

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By directly monitoring single cell growth in a microfluidic platform, we interrogated genome-deletion effects in Escherichia coli strains. We compared the growth dynamics of a wild type strain with a clean genome strain, and their derived mutants at the single-cell level. A decreased average growth rate and extended average lag time were found for the clean genome strain, compared to those of the wild type strain. Direct correlation between the growth rate and lag time of individual cells showed that the clean genome population was more heterogeneous. Cell culturability (the ratio of growing cells to the sum of growing and nongrowing cells) of the clean genome population was also lower. Interestingly, after the random mutations induced by a glucose starvation treatment, for the clean genome population mutants that had survived the competition of chemostat culture, each parameter markedly improved (i.e., the average growth rate and cell culturability increased, and the lag time and heterogeneity decreased). However, this effect was not seen in the wild type strain; the wild type mutants cultured in a chemostat retained a high diversity of growth phenotypes. These results suggest that quasi-essential genes that were deleted in the clean genome might be required to retain a diversity of growth characteristics at the individual cell level under environmental stress. These observations highlight that single-cell microfluidics can reveal subtle individual cellular responses, enabling in-depth understanding of the population.

Item Type:Articles
Keywords:Cell growth, genome reduction, microfluidics, mutation, single-cell.
Glasgow Author(s) Enlighten ID:Yin, Professor Huabing and Song, Dr Yanqing and Sloan, Professor William and Couto, Dr Jillian and Yuan, Dr Xiaofei and Glidle, Dr Andrew
Authors: Yuan, X., Couto, J. M., Glidle, A., Song, Y., Sloan, W., and Yin, H.
College/School:College of Science and Engineering > School of Engineering > Biomedical Engineering
College of Science and Engineering > School of Engineering > Infrastructure and Environment
Journal Name:ACS Synthetic Biology
Publisher:American Chemical Society
ISSN (Online):2161-5063
Published Online:26 August 2017
Copyright Holders:Copyright © 2017 American Chemical Society
First Published:First published in ACS Synthetic Biology 6(12): 2219-2227
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
621351Synthetic Biology applications to Water Supply and RemediationSteven BeaumontEngineering and Physical Sciences Research Council (EPSRC)EP/K038885/1VPO VICE PRINCIPAL RESEARCH & ENTERPRISE