Studies of complicated systems of growth using GH transgenic Amago
| Project/Area Number |
14360104
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General fisheries
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| Research Institution | Nihon University |
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Principal Investigator |
MORI Tsukasa Nihon University, Department of Nihon University College of Bioresource Sciences, Lecturer, 生物資源科学部, 講師 (60241379)
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| Co-Investigator(Kenkyū-buntansha) |
NAGOYA Hiroyuki Inland Station, National Research Institute of Aquaculture, 水産総合研究センター・養殖研究所・遺伝育種部, 主任研究員
海谷 啓之 国立循環器病センター, 研究所, 研究員 (40300975)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥14,700,000 (Direct Cost: ¥14,700,000)
Fiscal Year 2004: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2003: ¥9,000,000 (Direct Cost: ¥9,000,000)
Fiscal Year 2002: ¥3,600,000 (Direct Cost: ¥3,600,000)
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| Keywords | GH transgenic Amago / subtraction / microarray / DIGE / 肝臓、腸、脳下垂体 / F2,F3 / GHトランスジエニックアマゴ / Ghrelin遺伝子 / GHトランスジェニックアマゴF1 / ニジマスGHS-R遺伝子 / Ghrelin遺伝子組み換えニジマス |
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| Research Abstract |
(1)Proteins analysis extracted from intestine, liver and serum using DIGE
Proteins were extracted from intestine, liver and serum of GH transgenic Amago, and these proteins were performed proteins analysis using DIGE. About 2000 proteins were separated by two-dimensional gel electro-phoresisis, and some of major proteins were extracted from the gel and performed MS/MS-analysis. Most of these were unknown proteins, however, amount of digestive enzymes and G-proteins were significantly greater in transgenic intestine than that of control. Analysis of liver tissue also showed vast differences among transgenic and control. There were proteins having similar to parasites amino sequences in transgenic, and these were about four times grater than control. These proteins were detected only all transgenic used for the experiment.
(2)Analysis gene expression by microarray
We have performed subtractive hybridization using liver tissue from GH transgenic and control, and made microarray plates using these subtracted DNA. Analysis of microarray was performed using RNA extracted from both F2 and F3 of immature and mature GH transgenic. The results of these experiments showed drastic decrease of natural immune system in immature young fish. However, this tendency was getting installed when they were matured. Further, some sex related gene expression was observed in immature transgenic, and these tendency were also showed both F2 and F3.
(3)Screening of pituitary genes by subtraction
The completed length of cDNA library was obtained using pituitary of immature GH transgenic fish (F2), and also subtractive hybridization was performed using immature GH transgenic. Each 1000 clones of subtracted genes from both higher expression and lower expression were screened, and these have been sequenced.
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Report
(4 results)
Research Products
(2 results)
