Project Number: 3091-22000-031-29
Start Date: Jan 01, 2014
End Date: Dec 31, 2014
FOV strains will be grown in the lab under conditions that induce FA synthesis. mRNA will be extracted from cultures at times of rapidly increasing FA synthesis. Degenerate primers will be prepared to correspond with the conserved region of the ketoacyl synthases (KS) domain of the polyketide synthase (PKS) genes. Gene fragments of expected lengths will be excised, cloned, and transferred into Escherichia coli cells using appropriate vectors. Plasmids will be isolated from the transformed cells and the insert sequenced. The cloned PKS gene fragments will be identified using BLAST searches. Once PKS genes are identified by sequence homology, primers will be synthesized, and mRNA from cultures will be used as a template for polymerase chain reaction (PCR) analysis of the expression profile.