Using an AraC-based three-hybrid system to detect biocatalysts in vivo View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2000-05

AUTHORS

Steven M Firestine, Frank Salinas, Andrew E Nixon, Stephen J Baker, Stephen J Benkovic

ABSTRACT

Recent methods to create large libraries of proteins have greatly advanced the discovery of proteins with novel functions. However, one limitation in the discovery of new biocatalysts is the screening or selection methods employed to find enzymes from these libraries. We have developed a potentially general method termed QUEST (QUerying for EnzymeS using the Three-hybrid system), which allows the construction of an easily screened or selected phenotype for, in theory, any type of enzymatic reaction. The method couples the in vivo concentration of an enzyme's substrate to changes in the transcriptional level of a reporter operon. Using the arabinose operon activator AraC, we constructed a system capable of detecting the fungal enzyme scytalone dehydratase (SD) in bacteria, and demonstrated its sensitivity and usefulness in library screening. More... »

PAGES

nbt0500_544

Journal

TITLE

Nature Biotechnology

ISSUE

5

VOLUME

18

Author Affiliations

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/75414

    DOI

    http://dx.doi.org/10.1038/75414

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1023547905

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/10802623


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