| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2005: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2004: ¥2,500,000 (Direct Cost: ¥2,500,000)
|
|---|
| Research Abstract |
In this study, the petal cell maturing process of Torenia plant was investigated by using laser single-cell sampling and analysis technique. In the first year, sample preparation method and laser irradiation condition, and nano-flow LC-MS analysis condition of single-cell origin samples were investigated. Through this investigation, the facile analytical method for petal cell pigment was established. In second year, the quantitative analysis of mRNA concerning petal pigment biosynthesis was investigated. In particular, novel high sensitive quantitative mRNA analysis based on the ESI-MS was developed. In this method, the reporter molecule such as FAM (fluorescein phosphoramidite) was attached to the 5' end of the PCR primer and according to the PCR polymerizing reaction, the reporter molecule and the 5'end nucleotide was released. By quantify the released molecule using ESI-MS, the amount of targeted mRNA in the sample was estimated. The FAM attached nucleotide (FAM-A, FAM-T, FAM-G, FAM-C) was completely separated and quantified by LC-ESI-MS. Using this method, we are succeeded in the estimation of CHS and GADPH quantities in single petal cell of Torenia plant.
|
