In contrast to that of Anabaena sp. PCC 7120, the fdxN gene in Anabaena variabilis ATCC 29413 is not interrupted by a 55-kb DNA element, making this strain more suitable for genetic analysis of fdxN independent of the developmentally regulated excision during heterocyst formation. As a basis for mutational analysis, the fdxN gene of A. variabilis was cloned and sequenced. The deduced FdxN protein sequence was highly homologous to the Anabaena 7120 fdxN gene product including eight cysteine residues that are known to be conserved among ferredoxins containing two [4Fe-4S] clusters. The fdxN gene of A. variabilis was disrupted by insertion of an interposon within the fdxN coding region resulting in mutant strain KG29. Diazotrophic growth and in vivo nitrogenase activity of KG29 were similar to those of the wild-type, indicating that FdxN was not essential for N2 fixation in A. variabilis.