Oxygen Sensitivity of NifA Protein of Azospirillum lipoferum FS as Suggested by Gene Cloning and Expression in Escherichia coli.

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  • Oxygen Sensitivity of NifA Protein of A
  • Oxygen Sensitivity of NifA Protein of <i>Azospirillum lipoferum</i> FS as Suggested by Gene Cloning and Expression in <i>Escherichia coli</i>

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We cloned and sequenced a 2.8-kb SalI fragment of Azospirillum lipoferum FS as a homologue of the Klebsiella oxytoca nifA gene. The amino acid sequence deduced from an open reading frame of 1872 bases showed 91% identity to that of the A. brasilense NifA, and the putative central σ54 interaction domain was conserved as well as the C-terminal DNA-binding domain. The NifA function on the nifH promoter was examined in Escherichia coli using a combination of a nifA driver plasmid and a nifH-lacZ reporter plasmid, in which the transcriptional activation of the nifH promoter by the NifA was evaluated with the β-galactosidase activity. The A. lipoferum NifA activated the nifH promoter solely under microaerobic conditions, while the K oxytoca NifA activated it irrespective of the oxygen condition. These observations suggest that oxygen sensitivity is an intrinsic property of the A. lipoferum NifA.

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