Strongly enhanced bacterial bioluminescence with the ilux operon for single-cell imaging

Significance The emission of light generated in a process referred to as bioluminescence can be used for imaging of living cells over long timespans without phototoxicity or bleaching. The amounts of light produced in the bioluminescence process are very low, and exogenous substrate molecules are often required. We improved the brightness of bacterial bioluminescence, a system that features the advantage that all of the required molecular components are genetically encoded within a single operon. Consequently, we have engineered an improved operon ilux, which enables long-term visualization of single bacterial cells while simultaneously providing information about cellular viability. 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.

• Publication year

  2018

• Venue

  Proceedings of the National Academy of Sciences of the United States of America

• Publication date

  2018-01-16

• Fields of study

  Biology, Medicine, Chemistry, Engineering

• Identifiers
  DOI 10.1073/pnas.1715946115PMID 29339494PMCID 5798359
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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