Ratiometric Gas Reporting: A Nondisruptive Approach
To Monitor Gene Expression in Soils
Version 2 2018-02-20, 19:19
Version 1 2018-02-20, 19:12
Posted on 2018-02-20 - 19:19
Fluorescent proteins are ubiquitous
tools that are used to monitor
the dynamic functions of natural and synthetic genetic circuits. However,
these visual reporters can only be used in transparent settings, a
limitation that complicates nondisruptive measurements of gene expression
within many matrices, such as soils and sediments. We describe a new
ratiometric gas reporting method for nondisruptively monitoring gene
expression within hard-to-image environmental matrices. With this
approach, C2H4 is continuously synthesized by
ethylene forming enzyme to provide information on viable cell number,
and CH3Br is conditionally synthesized by placing a methyl
halide transferase gene under the control of a conditional promoter.
We show that ratiometric gas reporting enables the creation of Escherichia coli biosensors that report on acylhomoserine
lactone (AHL) autoinducers used for quorum sensing by Gram-negative
bacteria. Using these biosensors, we find that an agricultural soil
decreases the bioavailable concentration of a long-chain AHL up to
100-fold. We also demonstrate that these biosensors can be used in
soil to nondisruptively monitor AHLs synthesized by Rhizobium
leguminosarum and degraded by Bacillus thuringiensis. Finally, we show that this new reporting approach can be used in Shewanella oneidensis, a bacterium that lives in sediments.
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Cheng, Hsiao-Ying; Masiello, Caroline A.; Del Valle, Ilenne; Gao, Xiaodong; Bennett, George N.; Silberg, Jonathan J. (2018). Ratiometric Gas Reporting: A Nondisruptive Approach
To Monitor Gene Expression in Soils. ACS Publications. Collection. https://doi.org/10.1021/acssynbio.7b00405
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AUTHORS (6)
HC
Hsiao-Ying Cheng
CM
Caroline A. Masiello
ID
Ilenne Del Valle
XG
Xiaodong Gao
GB
George N. Bennett
JS
Jonathan J. Silberg