Development of Additional Selectable Markers for the Halophilic Archaeon<i>Haloferax volcanii</i>Based on the<i>leuB</i>and<i>trpA</i>Genes
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- Thorsten Allers
- Institute of Genetics, University of Nottingham, Queen's Medical Centre, Nottingham NG7 2UH, United Kingdom
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- Hien-Ping Ngo
- Institute of Genetics, University of Nottingham, Queen's Medical Centre, Nottingham NG7 2UH, United Kingdom
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- Moshe Mevarech
- Department of Molecular Microbiology and Biotechnology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel
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- Robert G. Lloyd
- Institute of Genetics, University of Nottingham, Queen's Medical Centre, Nottingham NG7 2UH, United Kingdom
説明
<jats:title>ABSTRACT</jats:title><jats:p>Since most archaea are extremophilic and difficult to cultivate, our current knowledge of their biology is confined largely to comparative genomics and biochemistry.<jats:italic>Haloferax volcanii</jats:italic>offers great promise as a model organism for archaeal genetics, but until now there has been a lack of a wide variety of selectable markers for this organism. We describe here isolation of<jats:italic>H. volcanii leuB</jats:italic>and<jats:italic>trpA</jats:italic>genes encoding 3-isopropylmalate dehydrogenase and tryptophan synthase, respectively, and development of these genes as a positive selection system. Δ<jats:italic>leuB</jats:italic>and Δ<jats:italic>trpA</jats:italic>mutants were constructed in a variety of genetic backgrounds and were shown to be auxotrophic for leucine and tryptophan, respectively. We constructed both integrative and replicative plasmids carrying the<jats:italic>leuB</jats:italic>or<jats:italic>trpA</jats:italic>gene under control of a constitutive promoter. The use of these selectable markers in deletion of the<jats:italic>lhr</jats:italic>gene of<jats:italic>H. volcanii</jats:italic>is described.</jats:p>
収録刊行物
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- Applied and Environmental Microbiology
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Applied and Environmental Microbiology 70 (2), 943-953, 2004-02
American Society for Microbiology
