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Achromobacter xylosoxidans KF701 and Pseudomonas putida (NAH7) were significantly different in degradative capability of aromatic compounds including bezoates, biphenyls, and naphthalene. Howeveer, both of the bacterial strains can grown on catechol as the sole carbon and energy source. Catechol 2,3-dioxygenase gene for naphthalene oxidation or biphenyl oxidation was cloned into Escherichia coli HB101 from NAH7 mega-plasmid of P. putida or chromosomal DNA of A. xylosoxidans KF701. A E. coli HB101 clone containting catechol 2,3-dioxygenase gene from P. putida (NAH7)contains a recombinant plasmid with 3.6kb pBR322 and 6-kb insert DNA. Physical maps of the recombinant plasmids were constructed, and catechol 2,3-dioxygenase gene in the recombinant plasmid was further localized and subcloned into M13. The cloned-catechol 2,3-dioxygenase gene products were identified as yellow bands on nondenaturaing polyaclamide gel after electrophoresis followed by activity staining with catechol solution.
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