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J Am Chem Soc. 2010 Sep 1;132(34):11902-3. doi: 10.1021/ja104550p.

Fluostatins produced by the heterologous expression of a TAR reassembled environmental DNA derived type II PKS gene cluster.

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Laboratory of Genetically Encoded Small Molecules, Howard Hughes Medical Institute, The Rockefeller University, New York, New York 10065, USA.


Culture independent approaches for accessing small molecules produced by uncultured bacteria are often hampered by the inability to easily clone environmental DNA (eDNA) fragments large enough to capture intact biosynthetic gene clusters that can be used in heterologous expression studies. Here we show that homology screening of eDNA megalibraries for clones containing natural product biosynthetic genes, coupled with transformation-assisted recombination (TAR) in yeast, can be used to access large, functionally intact, natural product gene clusters from the environment. The eDNA derived gene cluster reported here was functionally reconstructed from two overlapping cosmid clones using TAR. The isolation and structure elucidation of three new fluostatins (F, G, and H) produced by this TAR reconstructed gene cluster is described.

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