| Project/Area Number |
10556003
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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 | 展開研究 |
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| Research Field |
Breeding science
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| Research Institution | Natiomal Institute of Genetics |
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Principal Investigator |
KURATA Nori Natiomal Institute of Genetics, Genetic Strains Research Center, Associate Professor, 系統生物研究センター, 助教授 (90178088)
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| Co-Investigator(Kenkyū-buntansha) |
NONOMURA Kenichi Natiomal Institute of Genetics,Experimental Farm,Assistant Professor, 実験圃場, 助手 (10291890)
ITO Yukihiro Natiomal Institute of Genetics,Genetic Strains Research Center,Assistant Professor, 系統生物研究センター, 助手 (70280576)
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| Project Period (FY) |
1998 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥12,200,000 (Direct Cost: ¥12,200,000)
Fiscal Year 2001: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2000: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1999: ¥4,300,000 (Direct Cost: ¥4,300,000)
Fiscal Year 1998: ¥3,400,000 (Direct Cost: ¥3,400,000)
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| Keywords | キーワード無し / エンハンサートラップ / Ac / Ds / GUS遺伝子 / 転移系統 / Dsトランスポゾン / 形質転換体 / Ds-GUS転移 / 染色体挿入シリーズ / エンハンサースクリーニング |
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| Research Abstract |
(1) Generation of transgenic rice lines having single copy (42 lines) or double copies (above 50 lines) of Ds-GUS sequences and 6 lines possesing 35S-Ac transposase genes were generated. Examination for DsGUS transposition frequencies by screening F2 plants derived from cross combinations between four DsGUS and six 35S-Ac TPase lines with PCR method showed average frequency was 6% (675 / 10, 524 plants). (2) The 35S-Ac TPase line which showed higest activity to traspose Ds-GUS element was crossed with 22 Ds-GUS lines. A part ofF2 progenies, 926 out of 3277 F2 plants, exhibited transposition and average frequency of Ds-GUS transposition was calculated 28%. (3) We next screened 540 F2 transposants for GUS expression by staining almost all tissues in different developmental stages ; leaf blade, leaf sheath, auricle, ligule, root, node, rachis, papillae, style, ovary, anther, filament, lodicle, palea, lemma, upper glume, lower glume etc. Between 10 to 20 % of the transposants examined so far revealed to show trapped enahancer-GUS activity in any tissue. (4) To detect original and transposed insertion sites of Ds-GUS elememt, we isolated flanking genome sequences of the insertion positions and sequenced. A part of the flanking sequences (about 100) isolated and sequenced could be mapped through homology searching against chromosome assigned genome sequences. Remaining clones will be soon mapped on the fully aligned genome sequences
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