Development of a new method for the quantification of isoprenoid intermediates and analysis of the localization of the isoprenoid metabolic pathways in chloroplasts

Research Project

Project/Area Number	26650088
Research Category	Grant-in-Aid for Challenging Exploratory Research
Allocation Type	Multi-year Fund
Research Field	Plant molecular biology/Plant physiology
Research Institution	Hokkaido University
Principal Investigator	TANAKA Ryouichi 北海道大学, 低温科学研究所, 准教授 (20311516)
Project Period (FY)	2014-04-01 – 2015-03-31
Project Status	Completed (Fiscal Year 2014)
Budget Amount *help	¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000) Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Keywords	葉緑体
Outline of Final Research Achievements	We aimed at the development of a method to quantify geranylgeranyl-diphosphate (GGDP) and phytyl-diphosphate (PDP) in this study. First, we have tested whether geranylgeranyl transferase can conjugate GGDP and PDP to a dansylated peptide, which can be easily detected by a liquid chromatography system according to the method of Tong et al. 2013. However, this method only detect GGDP-conjugated peptide. Then, we attempted to use chlorophyll synthase activity to detect GGDP- or PDP-conjugate to chlorophyllide. However, we failed to overexpress this enzyme by an E. coli system, cultured insect cells (Sf9), and the MembraneMax cell-free expression system.

Report

(2 results)