Abstract
A glutamine synthetase gene ( glnA) was isolated from a deep-sea piezophilic bacterium, Shewanella violacea strain DSS12. A 7.5-kb SacI fragment containing the complete glnA gene was cloned and sequenced. The glnA gene was found to encode a protein consisting of 469 amino acid residues, showing 75.0% identity to the glutamine synthetase of Escherichia coli. Primer extension analyses revealed two transcription initiation sites in glnA and expression from each site was positively regulated by pressure. Putative promoters recognized by σ 70 and σ 54 were identified in the region upstream of glnA. An electrophoretic mobility shift assay demonstrated that S. violacea σ 54 specifically binds to the promoter region of glnA, suggesting that σ 54 may play an important role in pressure-regulated transcription in this piezophilic bacterium.
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