Heterologous expression in Saccharomyces cerevisiae of an Arabidopsis thaliana cDNA encoding mevalonate diphosphate decarboxylase.

摘要: Sequence comparison with the mevalonate diphosphate decarboxylase (MVD) amino acid sequence of Saccharomyces cerevisiae identified an EST clone corresponding to a cDNA that may encode Arabidopsis thaliana MVD (AtMVD1). This enzyme catalyses synthesis isopentenyl diphosphate, building block sterol and isoprenoid biosynthesis, uses as substrate. Sequencing full-length was performed. The predicted presents about 55% identity yeast, human rat MVDs. genomic region A. also obtained Southern blot analysis on DNA showed could have at least one homologous gene. In order allow heterologous expression in S. cerevisiae, open reading frame (ORF) then cloned under control yeast PMA1 strong promoter. When expressed complemented both thermosensitive MN19-34 strain deficient MVD, lethal phenotype ERG19 deleted strain. However, wild-type content not fully restored suggesting activity be optimal yeast. A two-hybrid assay performed evaluate homodimer formation heterodimer between plant enzymes.