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Applied and Environmental Microbiology, December 2006, p. 7485-7494, Vol. 72, No. 12
0099-2240/06/$08.00+0     doi:10.1128/AEM.01503-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Reconstitution of the Myxothiazol Biosynthetic Gene Cluster by Red/ET Recombination and Heterologous Expression in Myxococcus xanthus

Institut für Pharmazeutische Biotechnologie, Universität des Saarlandes, Postfach 15 11 50, D-66041 Saarbrücken, Germany,¹ Gene Bridges GmbH, Tatzberg 47-49, 01307 Dresden, Germany,² BioInnovationsZentrum, Department of Genomics, Technische Universität Dresden Tatzberg 47-51, 01307 Dresden, Germany³

Received 29 June 2006/ Accepted 18 September 2006

Although many secondary metabolites exhibiting important pharmaceutical and agrochemical activities have been isolated from myxobacteria, most of these microorganisms remain difficult to handle genetically. To utilize their metabolic potential, heterologous expression methodologies are currently being developed. Here, the Red/ET recombination technology was used to perform all required gene cluster engineering steps in Escherichia coli prior to the transfer into the chromosome of the heterologous host. We describe the integration of the complete 57-kbp myxothiazol biosynthetic gene cluster reconstituted from two cosmids from a cosmid library of the myxobacterium Stigmatella aurantiaca DW4-3/1 into the chromosome of the thus far best-characterized myxobacterium, Myxococcus xanthus, in one step. The successful integration and expression of the myxothiazol biosynthetic genes in M. xanthus results in the production of myxothiazol in yields comparable to the natural producer strain.

Published ahead of print on 22 September 2006.

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