| Budget Amount *help |
¥3,930,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥330,000)
Fiscal Year 2007: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2006: ¥2,500,000 (Direct Cost: ¥2,500,000)
|
|---|
| Research Abstract |
When I started this project,I planned to make a transgenic medaka which shows responsibility to chemicals such as dioxin utilizing GFP gene from jellyfish and the regulatory regions of fish CYP1 genes such as CYP1B1 and CYP1C genes from eel or carp,to verify the obtained responsibility to chemicals,and to select transgenic medaka which shows rapid response with intense fluorescence in gills. I could successfully make transgenic medaka using eel CYP1B1 gene. The obtained medaka exhibited intense GFP fluorescence comparable to that of CYP1A medaka in intestine,liver,and gills when induced by naphthoflavone as low as 500 ppb. The fluorescence intension in gills was clearly visible. Real time PCR study with the DNA fragments from the regulatory region of CYP1B1 gene,3.2 kbp,2.2 kbp,and 1.6 kbp,showed highest inductivity with 3.2 kbp fragment and lowest inductivity with 1.6 kbp fragment,revealing that XRE and ERE-like sequences play important roles in the induction.Unfortunately,F2 obtained by crossing the transgenic medaka (F1) with wild medaka was rather week to breed in the laboratory. Artificial construction of regulatory sequences which shows much higher inductivity may allow us to reduce the amount of DNA injected to a medaka egg and to produce transgenic medaka with more viability.
|
