Switchable gene expression in Escherichia coli using a miniaturized photobioreactor.
We present a light-switchable gene expression system for both inducible and switchable control of gene expression at a single cell level in Escherichia coli using a previously constructed light-sensing system. The λ cI repressor gene with an LVA degradation tag was expressed under the control of the ompC promoter on the chromosome. The green fluorescent protein (GFP) gene fused to a λ repressor-repressible promoter was used as a reporter. This light-switchable system allows rapid and reversible induction or repression of expression of the target gene at any desired time. This system also ensures homogenous expression across the entire cell population. We also report the design of a miniaturized photobioreactor to be used in combination with the light-switchable gene expression system. The miniaturized photobioreactor helps to reduce unintended induction of the light receptor due to environmental disturbances and allows precise control over the duration of induction. This system would be a good tool for switchable, homogenous, strong, and highly regulatable expression of target genes over a wide range of induction times. Hence, it could be applied to study gene function, optimize metabolic pathways, and control biological systems both spatially and temporally.
School of Nano-Bioscience and Chemical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan, Republic of Korea.
Jae Myung
JM
Lee
Junhyeong
J
Lee
Taesung
T
Kim
Sung Kuk
SK
Lee
D004926:Escherichia coli / Q000166:cytology / Q000235:genetics* / Q000528:radiation effects*; D015870:Gene Expression / Q000528:radiation effects; D005818:Genetic Engineering / Q000295:instrumentation / Q000379:methods*; D008027:Light; D008904:Miniaturization; D058433:Photobioreactors / Q000382:microbiology*; D059010:Single-Cell Analysis