Development of Additional Selectable Markers for the Halophilic ArchaeonHaloferax volcaniiBased on theleuBandtrpAGenes

Top Cited Papers
Publisher Website
Google Scholar
Abstract
Since most archaea are extremophilic and difficult to cultivate, our current knowledge of their biology is confined largely to comparative genomics and biochemistry.Haloferax volcaniioffers 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 ofH. volcanii leuBandtrpAgenes encoding 3-isopropylmalate dehydrogenase and tryptophan synthase, respectively, and development of these genes as a positive selection system. ΔleuBand ΔtrpAmutants 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 theleuBortrpAgene under control of a constitutive promoter. The use of these selectable markers in deletion of thelhrgene ofH. volcaniiis described.