关键词: 青钱柳, 法呢基焦磷酸合成酶, RACE, 功能研究

Abstract: Cyclocarya paliurus is a medicinal, timber and ornamental tree. Farnesyl diphosphate synthase (FPS; EC2.5.1.1/EC2.5.1.10) catalyses 1′-4 condensation of the 5-carbon isoprenoid compounds isopentenyl diphosphate (IPP) and 10-carbon geranyl diphosphate (GPP) to form the 15-carbon product farnesyl diphosphate (FPP). FPP is the common precursor of sesquiterpenes, tritepenes, steroids, etc. cDNA cloning and characterization of a novel FPS from C.paliurus were described in this present study. The full-length cDNA named CpFPS(Genbank Accession Number GU121224) contained 1 420 bp with an open reading frame of 1 029 bp encoding a polypeptide of 342 amino acids with a calculated molecular mass of 39.60 kDa. Through sequence analysis by BlASTP online, the resulting protein showed high homology to the FPS of Gossypium arboretum(CAA72793) and Hevea brasiliensis(BAF98301). Deduced amino acid sequence was also analyzed by biological software and the data presented the existence of conserved and functional domains of FPS. The functional analysis of CpFPS was performed by using the sterol-auxotrophic yeast strain CC25. The result indicated that the cloned cDNA of CpFPS encoding a functional FPS in C.paliurus.

Key words: Cyclocarya paliurus, farnesyl diphosphate synthase(FPS), RACE, functional analysis