Summary: The surface-adhering, Gram-negative marine bacterium synthesizes a red-brown melanin in the late stage of exponential growth in laboratory culture. Previous studies identified a single gene, , from that could impart the ability to produce melanin to an host. However, these studies did not demonstrate a requirement for during melanization in In this paper, genetic analyses, using a broad host range conjugation system to generate specific lesions, reveal that null mutants fail to synthesize pigment. The wild-type gene provided on a low copy number plasmid complemented these null mutations, as well as a spontaneous pigment variant, to wild-type melanin synthesis. Polyclonal antibodies, raised against a MelA-LacZ fusion protein, were used to confirm the presence of the gene product in wild-type and verify its absence in the non-pigmented mutants. In addition, detection of the MelA protein over the course of growth in batch culture revealed a constant steady-state level of MelA protein, suggesting that the timing of melanizarion and the quantity of melanin synthesized is not controlled at the level of expression.


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