| Δ3-Δ2-Enoyl-CoA isomerase from the yeast Saccharomyces cerevisiae: Molecular and structural characterization | ||
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The ECI1 knockout yeast strain was constructed by the short flanking homology method (Wach et al. 1994) based on polymerase chain reaction (PCR) -targeting following the European Functional Analysis Network guidelines. In this method, the target gene is replaced by a kanMX cassette, which gives the knockout strain resistance to the drug geneticin. To test the ability of the ECI1-disrupted strain to metabolize saturated and unsaturated fatty acids, the mutant strain was grown on palmitic (C16:0), oleic [cis-C18:1(9)], octadecenoic [cis-C18:1(12)] or arachidonic acid [cis-C20:4(5, 8, 11, 14)] as the sole carbon source. The experiments were made in the Institute of Biochemistry and Molecular Cell Biology, University of Vienna, Austria.