Activity-Based Screening of Metagenomic Libraries for Hydrogenase Enzymes.

Adam, Nicole and Perner, Mirjam (2017) Activity-Based Screening of Metagenomic Libraries for Hydrogenase Enzymes. In: Metagenomics: Methods and Protocols. , ed. by Streit, Wolfgang R. and Daniel, Rolf. Springer, New York, NY, pp. 261-270. ISBN 978-1-4939-6689-9 DOI 10.1007/978-1-4939-6691-2_17.

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Abstract

Here we outline how to identify hydrogenase enzymes from metagenomic libraries through an activity-based screening approach. A metagenomic fosmid library is constructed in E. coli and the fosmids are transferred into a hydrogenase deletion mutant of Shewanella oneidensis (ΔhyaB) via triparental mating. If a fosmid exhibits hydrogen uptake activity, S. oneidensis' phenotype is restored and hydrogenase activity is indicated by a color change of the medium from yellow to colorless. This new method enables screening of 48 metagenomic fosmid clones in parallel.

Document Type: Book chapter
Keywords: Metagenome, Function-based screen, Hydrogenase, Hydrogen uptake
Open Access Journal?: No
DOI etc.: 10.1007/978-1-4939-6691-2_17
Date Deposited: 13 Nov 2018 08:02
Last Modified: 14 Dec 2018 10:55
URI: http://oceanrep.geomar.de/id/eprint/44629

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