Mutants of ADP1 unable to grow on dodecane, but retaining the ability to grow on lauric acid were isolated after ethylmethanesulphonate (EMS) treatment. This growth deficiency was complemented by a clone from a gene library constructed from chromosomal DNA of the wild-type strain. The complementing DNA mapped in a gene encoding a polypeptide with homology to rubredoxins. The deduced putative rubredoxin amino acid sequence is more similar to related proteins from Gram-positive bacteria than to the rubredoxin involved in alkane oxidation. An adjacent gene encodes a protein with similarity to rubredoxin reductase from and related NAD(P)-dependent reductases. Disruption of the rubredoxin-encoding gene by insertion of a Km/ cassette rendered the resulting strain unable to grow on dodecane or hexadecane. This demonstrates that these genes are necessary for alkane degradation. Transcriptional fusion of to the rubredoxin-encoding gene led to low level constitutive -galactosidase expression, whereas the fusion oriented in the opposite direction was not expressed.


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