cDNA Cloning and Expression Analysis of Farnesyl Pyrophosphate Synthase from Ornithogalum saundersiae
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Lei Guo
Farnesyl pyrophosphate synthase (FPPS, EC 2.5.1.10) catalyzes the consecutive head-to-tail condensations of isopentenyl diphosphate (IPP) with dimethylallyl diphosphate (DMAPP) to form farnesyl pyrophosphate (FPP), a key precursor of sesquiterpenoids, triterpenoids, sterols, and farnesylated proteins. Here we report the molecular cloning and functional identification of a new full-length cDNA encoding FPPS from Ornithogalum saundersiae, a potential medicinal plant that produces a promising antitumour sterol glycoside, OSW-1. An 1327 bp long unigene with an open reading frame of 1044 bp was retrieved from the transcriptome sequencing of O. saundersiae. The full-length FPPS cDNA, designated OsaFPPS, was isolated from O. saundersiae with gene-specific primers. The resultant OsaFPPS encodes a 347-amino acids protein with a calculated molecular mass of 40;085:6 Da, and a theoretical isoelectric point of 5.01. Phylogenetic tree analysis indicated that OsaFPPS belongs to the plant FPPS super-family. Expression of soluble OsaFPPS in E. coli was verified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis. Functional analysis of the purified OsaFPPS protein was carried out using IPP and DMAPP as substrates, and the product was unambiguously determined by gas chromatography-mass spectrometry (GC-MS) analyses
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Articles in the same Issue
- Chemical Constituents of Croton oligandrum (Euphorbiaceae)
- Acacetin-7-rutinoside from Buddleja lindleyana, a New Molluscicidal Agent against Oncomelania hupensis
- Cytotoxic Saponins from the Seeds of Pittosporum angustifolium
- Flavone Composition and Antihypercholesterolemic and Antihyperglycemic Activities of Chrysanthemum coronarium L.
- New Nitrogenous Compounds from Anisotes trisulcus
- Inhibitory Effect of the Methanolic Extract of Verbascum latisepalum Hub.-Mor. on Endothelium-Dependent Relaxation in Rat Thoracic Aorta
- Exogenous Ascorbic Acid and Glutathione Alleviate Oxidative Stress Induced by Salt Stress in the Chloroplasts of Oryza sativa L.
- Improved ß-Carotene and Lycopene Production by Blakeslea trispora with Ultrasonic Treatment in Submerged Fermentation
- Microbial Metabolism of Danazol: A Contribution to Doping Analysis
- Identification of Two Chemotypes of Pogostemon cablin (Blanco) Benth. through DNA Barcodes
- cDNA Cloning and Expression Analysis of Farnesyl Pyrophosphate Synthase from Ornithogalum saundersiae
