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  • 标题:Strongly enhanced bacterial bioluminescence with the ilux operon for single-cell imaging
  • 本地全文:下载
  • 作者:Carola Gregor ; Klaus C. Gwosch ; Steffen J. Sahl
  • 期刊名称:Proceedings of the National Academy of Sciences
  • 印刷版ISSN:0027-8424
  • 电子版ISSN:1091-6490
  • 出版年度:2018
  • 卷号:115
  • 期号:5
  • 页码:962-967
  • DOI:10.1073/pnas.1715946115
  • 语种:English
  • 出版社:The National Academy of Sciences of the United States of America
  • 摘要:Bioluminescence imaging of single cells is often complicated by the requirement of exogenous luciferins that can be poorly cell-permeable or produce high background signal. Bacterial bioluminescence is unique in that it uses reduced flavin mononucleotide as a luciferin, which is abundant in all cells, making this system purely genetically encodable by the lux operon. Unfortunately, the use of bacterial bioluminescence has been limited by its low brightness compared with other luciferases. Here, we report the generation of an improved lux operon named ilux with an approximately sevenfold increased brightness when expressed in Escherichia coli ; ilux can be used to image single E. coli cells with enhanced spatiotemporal resolution over several days. In addition, since only metabolically active cells produce bioluminescent signal, we show that ilux can be used to observe the effect of different antibiotics on cell viability on the single-cell level.
  • 关键词:bioluminescence ; luciferase ; antibiotics ; bacteria ; microscopy
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