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Mol. Cells 2005; 19(2): 294-299
Published online January 1, 1970
© The Korean Society for Molecular and Cellular Biology
Cloning and Expression of a Farnesyl Diphosphate Synthase in Centella asiatica (L.) Urban
A cDNA encoding farnesyl diphosphate synthase (FPS; EC2.5.1.1/EC2.5.1.10) was isolated from Centella asiacita (L.) Urban, using degenerate primers based on two highly conserved domains. A full-length cDNA clone was subsequently isolated by rapid amplification of cDNA ends (RACE) PCR. The sequence of the CaFPS (C. asiatica farnesyl diphosphate synthase) cDNA contains an open reading frame of 1029 nucleotides encoding 343 amino acids with a molecular mass of 39.6 kDa. The deduced CaFPS amino acid sequence exhibits 84, 79, and 72%, identity to the FPSs of Artemisia annua, Arabidopsis thaliana, and Oryza sativa, respectively. Southern blot analysis suggested that the C. asiatica genome contains only one FPS gene. An artificially expressed soluble form of the CaFPS was identified by SDS-PAGE. It had high specific activity and produced farnesyl diphosphate as the major isoprenoid.
Keywords Centella asiatica; Farnesyl Diphosphate Synthase; Isoprenoid Pathway; Triterpenoid Saponins
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Communication
Mol. Cells 2005; 19(2): 294-299
Published online April 30, 2005
Copyright © The Korean Society for Molecular and Cellular Biology.
Cloning and Expression of a Farnesyl Diphosphate Synthase in Centella asiatica (L.) Urban
Ok Tae Kim, Jun Cheul Ahn, Sung Jin Hwang, Baik Hwang
Abstract
A cDNA encoding farnesyl diphosphate synthase (FPS; EC2.5.1.1/EC2.5.1.10) was isolated from Centella asiacita (L.) Urban, using degenerate primers based on two highly conserved domains. A full-length cDNA clone was subsequently isolated by rapid amplification of cDNA ends (RACE) PCR. The sequence of the CaFPS (C. asiatica farnesyl diphosphate synthase) cDNA contains an open reading frame of 1029 nucleotides encoding 343 amino acids with a molecular mass of 39.6 kDa. The deduced CaFPS amino acid sequence exhibits 84, 79, and 72%, identity to the FPSs of Artemisia annua, Arabidopsis thaliana, and Oryza sativa, respectively. Southern blot analysis suggested that the C. asiatica genome contains only one FPS gene. An artificially expressed soluble form of the CaFPS was identified by SDS-PAGE. It had high specific activity and produced farnesyl diphosphate as the major isoprenoid.
Keywords: Centella asiatica, Farnesyl Diphosphate Synthase, Isoprenoid Pathway, Triterpenoid Saponins
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