An easy cloning and expression vector system for Gluconobacter oxydans.

Gluconobacter oxydans is known for causing rapid and incomplete oxidation of a wide range of sugars, sugar acids and sugar alcohols. Therefore, this microorganism is already employed in several biotechnological processes that involve incomplete oxidation of a substrate, e.g. vitamin C or dihydroxyacetone production. To fully exploit the oxidative potential of G. oxydans, characterization of the biological role of gene products is essential. To take advantage of the genome sequence of G. oxydans DSM 2343, based on pBBR1MCS5, we constructed a new cloning and expression vector. The newly established vector pEXGOX will significantly decrease duration of cloning and increase cloning efficiency. It has the following advantages: (i) small size (5.7 kbp); (ii) complete sequence; (iii) variety of unique restriction sites; (iv) direct cloning of PCR products; (v) strong promoter. The pEXGOX plasmid was successfully used to clone G. oxydans genes and has the potential to facilitate studies of gene function of several G. oxydans open reading frames.