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Media type:
E-Article
Title:
Towards a Systems Approach in the Genetic Analysis of Archaea: Accelerating Mutant Construction and Phenotypic Analysis inHaloferax volcanii
Contributor:
Blaby, Ian K.;
Phillips, Gabriela;
Blaby-Haas, Crysten E.;
Gulig, Kevin S.;
El Yacoubi, Basma;
de Crécy-Lagard, Valérie
Published:
Hindawi Limited, 2010
Published in:
Archaea, 2010 (2010), Seite 1-11
Language:
English
DOI:
10.1155/2010/426239
ISSN:
1472-3654;
1472-3646
Origination:
Footnote:
Description:
With the availability of a genome sequence and increasingly sophisticated genetic tools,Haloferax volcaniiis becoming a model for both Archaea and halophiles. In order forH. volcaniito reach a status equivalent toEscherichia coli, Bacillus subtilis, orSaccharomyces cerevisiae, a gene knockout collection needs to be constructed in order to identify the archaeal essential gene set and enable systematic phenotype screens. A streamlined gene-deletion protocol adapted for potential automation was implemented and used to generate 22H. volcaniideletion strains and identify several potentially essential genes. These gene deletion mutants, generated in this and previous studies, were then analyzed in a high-throughput fashion to measure growth rates in different media and temperature conditions. We conclude that these high-throughput methods are suitable for a rapid investigation of anH. volcaniimutant library and suggest that they should form the basis of a larger genome-wide experiment.