Fusarin C biosynthesis in Fusarium moniliforme and Fusarium venenatum.
Fragments of polyketide synthase (PKS) genes were amplified from complementary DNA (cDNA) of the fusarin C producing filamentous fungi Fusarium moniliforme and Fusarium venenatum by using degenerate oligonucleotides designed to select for fungal PKS C-methyltransferase (CMeT) domains. The PCR products, which were highly homologous to fragments of known fungal PKS CMeT domains, were used to probe cDNA and genomic DNA (gDNA) libraries of F. moniliforme and F. venenatum. A 4.0 kb cDNA clone from F. venenatum was isolated and used to prepare a hygromycin-resistance knockout cassette, which was used to produce a fusarin-deficient strain of F. venenatum (kb = 1000 bp). Similarly, a 26 kb genomic fragment, isolated on two overlapping clones from F. moniliforme, encoded a complete iterative Type I PKS fused to an unusual nonribosomal peptide synthase module. Once again, targeted gene disruption produced a fusarin-deficient strain, thereby proving that this synthase is responsible for the first steps of fusarin biosynthesis.
Song
Zhongshu
Z
School of Chemistry, University of Bristol, Cantock's Close, Clifton, Bristol, BS8 1TS, UK.
Cox
Russell J
RJ
Lazarus
Colin M
CM
Simpson TJ
Thomas J
TJ
D001224:Aspartic Acid / Q000737:chemistry / Q000378:metabolism; D015139:Blotting, Southern; D002853:Chromatography, Liquid; D002952:Citric Acid Cycle / Q000502:physiology; D017931:DNA Primers; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D005670:Fusarium / Q000254:growth & development / Q000378:metabolism*; D015723:Gene Library; D013058:Mass Spectrometry; D009153:Mutagens / Q000378:metabolism*; D062907:Oxaloacetic Acid / Q000737:chemistry / Q000378:metabolism; D010957:Plasmids; D011090:Polyenes / Q000737:chemistry / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012436:S-Adenosylmethionine / Q000737:chemistry / Q000378:metabolism; D014170:Transformation, Genetic / Q000187:drug effects
D017931:DNA Primers; D018076:DNA, Complementary; D009153:Mutagens; D011090:Polyenes; D012333:RNA, Messenger; D062907:Oxaloacetic Acid; D001224:Aspartic Acid; D011092:Polyethylene Glycols; D012436:S-Adenosylmethionine; C041403:fusarin C