Sunday, August 10, 2008
P101
A platform for studying of unknown PKS/NRPS gene clusters
Youming Zhang1, Jun Fu2, Fan Huang2, Shengbiao Hu2, Francis A. Stewart2, Thorsten Klefisch3, and Rolf Müller3. (1) R&D, Gene Bridges GmbH, BioZ, Tatzberg 47-51, Dresden, 01307, Germany, (2) Genomics, Biotec, Technology University of Dresden, BioZ, Tatzberg 47-51, Dresden, 01307, Germany, (3) Pharmaceutical Biotechnology, Saarland University, Saarbrücken, 66041, Germany
DNA sequencing points to a new way to discover secondary metabolites. Up to date, more than 600 microorganism genomes have been sequenced and there are more and more data coming from the Metagenomic sequencing project. Sequencing of two myxobacterial strains (Myxococcus xanthus and Sorangium cellulosum 56) reveals >20 clusters that are devoted to secondary metabolic pathways, a striking feature that is also common to the genomes of streptomycetes and mycobacteria. Complete genome sequencing has revealed that even well characterized Actinomycetes and Myxobacteria contain ‘silent’ or cryptic PKS/NRPS gene clusters. By sequence criteria, these gene clusters are functional however they do not express their secondary metabolite under the standard culture conditions. Genome sequencing does not identify new PKS/NRPS metabolites. It produces information and cloned DNA. Sophisticated DNA engineering to adapt cloned DNA pieces for expression in heterologous hosts is needed to evaluate the potential of novel compounds.
Homologous recombination mediated Recombineering is a unique technology for large size gene engineering and it has been applied for type I PKS/NRPS gene cluster engineering. Based on Recombineering and the newly developed transpositional technologies, one platform has been set up for exploring of the unknown gene clusters.
Homologous recombination mediated Recombineering is a unique technology for large size gene engineering and it has been applied for type I PKS/NRPS gene cluster engineering. Based on Recombineering and the newly developed transpositional technologies, one platform has been set up for exploring of the unknown gene clusters.
- Direct cloning of unknown gene clusters from microbial genomic DNA by Recombineering without library construction.
- Insertion of inducible or constitutive promoters in front of gene clusters.
- Insertion of functional cassettes into the vector backbone for mobilizing the gene clusters in different heterologous expression hosts.
- Transferring of gene clusters into heterologous hosts for expression.
- Compound detection and evaluation.
See more of Poster Session 1
See more of Posters
See more of The Annual Meeting and Exhibition 2008 (August 10 - 14, 2008)
See more of Posters
See more of The Annual Meeting and Exhibition 2008 (August 10 - 14, 2008)