Development of novel metabolite-responsive transcription factors via transposon-mediated protein fusion.

ABSTRACT: Naturally evolved metabolite-responsive biosensors enable applications in metabolic engineering, ranging from screening large genetic libraries to dynamically regulating biosynthetic pathways. However, there are many metabolites for which a natural biosensor does not exist. To address this need, we developed a general method for converting metabolite-binding proteins into metabolite-responsive transcription factors-Biosensor Engineering by Random Domain Insertion (BERDI). This approach takes advantage of an in vitro transposon insertion reaction to generate all possible insertions of a DNA-binding domain into a metabolite-binding protein, followed by fluorescence activated cell sorting to isolate functional biosensors. To develop and evaluate the BERDI method, we generated a library of can

SUBMITTER: Younger AKD 

PROVIDER: S-EPMC5914420 | biostudies-literature | 2018 Feb

REPOSITORIES: biostudies-literature