Submitted to: British Mycological Society Annual International Symposia Series
Publication Type: Abstract Only
Publication Acceptance Date: March 5, 2001
Publication Date: April 3, 2001
Citation: GIBSON, D.M., KRASNOFF, S.B., LEE, T., YUN, S., HODGE, K.T., HUMBER, R.A., TURGEON, G.B., YODER, O.C. USING A PCR-DIRECTED APPROACH TO NARROW THE SEARCH FOR BIOACTIVE METABOLITES FROM INSECT- AND NEMATODE-ASSOCIATED FUNGI. BRITISH MYCOLOGICAL SOCIETY ANNUAL INTERNATIONAL SYMPOSIA SERIES. 2001.
Production of polyketides is accomplished through complex enzymes known as polyketide synthases (PKS); these enzymes have highly conserved domains that might be useful in screens for PKSs in diverse groups of organisms. A degenerate PCR-based approach was used to amplify PKS fragments of the ketosynthase domain from genomic DNA of a group of insect- and nematode-associated fungi. Of 157 isolates (representing 73 genera and 14 species) screened, 92 isolates generated PCR products of predicted size (~ 300 bp). The ability to detect PKS domains was a function of the number of different primer pairs employed in the screen. Cloning and sequencing analysis revealed that 66 isolates had at least one unique PKS sequence; ten members of this set contained multiple PKS fragments, for a total of 76 unique PKS fragments. Since PKS genes appear to be widespread among fungi, a PCR-based screening system appears to be an efficient, directed means to identify organisms having the potential to produce polyketides.