A Biosensor Strategy for E. coli Based on Ligand-Dependent Stabilization.
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ABSTRACT: The engineering of microorganisms to monitor environmental chemicals or to produce desirable bioproducts is often reliant on the availability of a suitable biosensor. However, the conversion of a ligand-binding protein into a biosensor has been difficult. Here, we report a general strategy for generating biosensors in Escherichia coli that act by ligand-dependent stabilization of a transcriptional activator and mediate ligand concentration-dependent expression of a reporter gene. We constructed such a biosensor by using the lac repressor, LacI, as the ligand-binding domain and fusing it to the Zif268 DNA-binding domain and RNA polymerase omega subunit transcription-activating domain. Using error-prone PCR mutagenesis of lacI and selection, we identified a biosensor with multiple mutations,
SUBMITTER: Brandsen BM
PROVIDER: S-EPMC6358018 | biostudies-literature | 2018 Sep
REPOSITORIES: biostudies-literature
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