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| Codon Optimization | The open reading frames of DES1 and DES2 were codon optimised for expression in S. cerevisiae by Genscript, synthesised and supplied in the pUC-57 vector. DES1 and DES2 were then excised using BamHI and SalI restriction sites and ligated into pHEY1 [22], for expression under the constitutive TEF1 promoter. pHEY-DES1, pHEY-DES2 and pHEY-EVC (empty vector) were transformed into S. cerevisiae [23] wild type strain DTY-11a (MATa, leu2-3, leu2-12, trp1–1, can1–100, ura3–1, ade2–1) and ole1Δ knockout strain AMY-3α (MATα, ole1Δ::LEU2, trp1–1, can1–100, ura3–1, ade2–1) [24] and colonies were selected on synthetic Dexterose (SD) minimal medium agar plates lacking uracil. | Get A Quote |
Arbuscular mycorrhizal fungi are oleaginous organisms and the most abundant fatty acyl moiety usually found in their lipids is palmitvaccenic acid (16:1Δ11cis). However, it is not known how this uncommon fatty acid species is made. Here we have cloned two homologs of Lepidopteran fatty acyl-CoenzymeA Δ11 desaturases from Rhizophagus irregularis. Both DES1 and DES2 are expressed in intraradicle mycelium and can complement the unsaturated fatty acid-requiring auxotrophic growth phenotype of the Saccharomyces cerevisiae ole1Δ mutant. DES1 expression leads almost exclusively to oleic acid (18:1Δ9cis) production, whereas DES2 expression results in the production of 16:1Δ11cis and vaccenic acid (18:1Δ11... More
