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Functional identification of sterol‐4α‐methyl oxidase cDNAs from Arabidopsis thaliana by complementation of a yeast erg25 mutant lacking sterol‐4α‐methyl oxidation 1
Author(s) -
Darnet Sylvain,
Bard Martin,
Rahier Alain
Publication year - 2001
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(01)03002-2
Subject(s) - complementation , complementary dna , arabidopsis thaliana , biology , sterol , biochemistry , saccharomyces cerevisiae , ergosterol , cdna library , mutant , auxotrophy , heterologous expression , arabidopsis , yeast , microbiology and biotechnology , gene , recombinant dna , cholesterol
Specific primers derived from both genomic sequence data and EST cDNA sequences were used to polymerase chain reaction amplify two full‐length cDNA sequences ( At SMO1 and At SMO2), 801 and 783 bp, respectively, from an Arabidopsis thaliana cDNA library. The predicted proteins show 32 and 29% identity to the ERG25 gene from Saccharomyces cerevisiae which encodes the sterol‐4α‐methyl oxidase (SMO), a membrane‐bound non‐heme di‐iron oxygenase involved in lipid metabolism. Heterologous expression of At SMO1 and At SMO2 in a yeast erg25 ergosterol auxotroph, lacking SMO activity, restored growth and endogenous ergosterol synthesis. These results represent the first functional identification of SMO genes from plants.