Molecular cloning and functional analysis of an organ-specific expressing gene coding for farnesyl diphosphate synthase from Michelia chapensis Dandy

Abstract

Farnesyl diphosphate synthase (FPS; EC 2.5.1.1/EC 2.5.1.10) catalyzes the synthesis of farnesyl diphosphate, a key intermediate in the biosynthesis of sesquiterpenes. This present study described the cloning and characterization of a cDNA encoding FPS from leaves of Michelia chapensis Dandy (designated as McFPS, GenBank accession number: GQ214406) for the first time. McFPS was 1,432 bp and contained an open reading frame (ORF) of 1,056 bp, encoding a protein of 351 amino acids with a calculated molecular mass of 40.52 kDa. Bioinformatic analysis revealed that the deduced McFPS had high homology with FPSs from other plant species. Phylogenetic tree analysis indicated that McFPS belonged to the plant FPS group and had the closest relationship with FPS from Chimonanthus praecox. Southern blot analysis revealed that there were at most two copies of McFPS gene existed in M. chapensis genome. The organ expression pattern analysis showed that McFPS expressed strongly only in leaves, and there were no expression in stems and roots, implying that McFPS was an organ-specific expressing gene. Functional complementation of McFPS in a FPS-deficient yeast strain demonstrated that cloned cDNA encoded a farnesyl diphosphate synthase.