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In the study described here, we successfully developed a transformation system for halo(alkali)philic members of theArchaea . This transformation system comprises a series ofNatrialba magadii /Escherichia coli shuttle vectors based on a modified method to transform halophilic members of theArchaea and genomic elements of theN. magadii virus ϕCh1. The shuttle vector pRo-5, based on therepH -containing region of ϕCh1, stably replicated inE. coli andN. magadii and in several halophilic and haloalkaliphilic members of theArchaea not transformable so far. The ϕCh1 operon ORF53/ORF54 (repH ) was essential for pRo-5 replication and was thus identified as the minimal replication origin. The plasmid allowed homologous and heterologous gene expression, as exemplified by the expression of ϕCh1 ORF3452 , which encodes a structural protein, and the reporter genebgaH ofHaloferax lucentense inN. magadii . The new transformation/vector system will facilitate genetic studies withinN. magadii and other haloalkaliphilic archaea and will allow the detailed characterization of the gene functions ofN. magadii virus ϕCh1 in their extreme environments.

Utilization of Virus ϕCh1 Elements To Establish a Shuttle Vector System for Halo(alkali)philic Archaea via Transformation of Natrialba magadii