JP2010029196A - 遺伝子発現の特異性に関する改良 - Google Patents
遺伝子発現の特異性に関する改良 Download PDFInfo
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Abstract
【解決手段】植物の複数の領域において発現される植物内在性遺伝子の機能に影響を及ぼす作用物質に連結されているプロモーター。1以上の所望の位置にその発現部位におけるオーバーラップが存在するように選択されるプロモーターおよび作用物質。
【選択図】なし
Description
2.共同抑制
3.ターゲット遺伝子のプロモーターの阻害または活性化
4.転写の阻害または活性化
5.メッセンジャーRNA安定性の変更またはmRNAの分解
6.翻訳の阻害または活性化
7.タンパク質の阻害または活性化
8.タンパク質代謝回転の変更
9.共同因子としての作用
10.タンパク質−タンパク質相互作用の変更
11.生化学的経路によるフラックスの変更。
ヴェン構築物を用いた操作により植物寄生線虫に対する耐性または抵抗性を増大させた。
非形質転換対照植物、対照pBIN05101植物およびpBIN05002試験植物についての低えい瘤スコア区分と高えい瘤スコア区分における植物の数
低えい瘤スコア=0〜50えい瘤/植物
高えい瘤スコア=51+えい瘤/植物
処理 低えい瘤スコアの植物 高えい瘤スコアの植物
非形質転換植物 18 13
pBIN05101対照植物 13 17
pBIN05002系32試験植物 25 7
上記の実施例において、線虫破壊における特異性を増大させるための細胞破壊系を用いて説明したオーバーラップの原理は、例えば、Arabidopsis、またはその他の植物の花においても同様に十分に実行されて、花のパターンまたは構造が変化した花、例えば、雄性不稔性の花が得られ得る。この実施例は、Arabidopsisにおいて同定されたDNA配列を利用するものである。
プライマー1(配列番号1)
ATCGAAGCTT CTAAATGTAC TGAAAAGAAA CA
プライマー2(配列番号2)
ACTGGGATCC GAAAATGGAA GGTAAGGTTG TGC
プライマーは、ゲノムagamous配列についてのジーンバンクDNA配列エントリーATAGAMSGで得られた配列に基づくものであった。プライマー1はその5'末端に追加のHindIII制限部位を含む。プライマー2は、その5'末端に追加のBamHI制限部位を含む。
プライマー3(配列番号3)
ATCGGGATCC ATGGGCTCAC GGTTTTGTGT GA
プライマー4(配列番号4)
ATCGGAGCTC TTATTCAAGA AGATGGAAGG TAATGA
プライマー3は、公表されているapetala3配列の992位にて増幅が開始されるように特異的に設計され、これは活性産物を産生させるための不適正リーディングフレームにおける開始コドンであり、同一遺伝子ファミリー内のその他のMADSボックス遺伝子と強い相同性を有する配列の初期部分を無効にするものである。また、プライマー3は、望ましくないSacI制限部位を除去するために公表されている配列から1塩基対変更されている。プライマー3はその5'末端にBamHI制限部位を有している。プライマー4はその5'末端にSacI部位を有している。1586bp断片を、プライマー3および4を用いてPCRによってArabidopsis DNAから増幅させた。
プライマー7(配列番号7)
GAACTGGGAC CACTCCAGTG
いずれの場合においても、適切な構築物を含むトランスジェニック植物が同定された。
Bechtold, N., Ellis, J., およびPelletier, G. (1993) In planta Agrobacterium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants, C.R. Acad. Sci: Paris 316 : 1194-1199。
Bevan, M. (1984) Binary Agrobacterium vectors for plant transformation, Nuc. Acids Res. 12: 8711-8721。
Coen, E.S. およびMeyerowitz, E.M. (1991) The war of the whorls: genetic interactions controlling flower development, Nature 353, 31-37。
Conkling, M.A., Cheng, C-L., Yamamoto, Y.T. およびGoodman, H.M.(1990) Isolation of transcriptionally regulated root-specific genes from tobacco, Plant Physiology 93, 1203-1211。
Horsch, R.B., Fry, J.F., Hoffmann, N.L., Eichholtz, D., Rogers, S.G. およびFraley, R.T.(1985), A simple and general method for transferring genes into plants, Science 22, 1229-1231。
Jefferson, R.A., Kavanagh, T.A. およびBevan, M.W. (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion maker in higher plants, EMBO, 6, 3901-3907。
Opperman, C.H., Taylor, C.G. およびConkling, M.A. (1994) Root-knot nematode-directed expression of a plant root specific gene, Science 263, 221-223。
Sambrook, J., Fritsch, E.F. およびManiatis, T. (1989) Molecular Cloning - A Laboratory Manual. Second Edition, Cold Spring Harbour Laboratory Press, New York。
Thomas, C. (1996) 'PCR Techniques' in 'Plant Gene Isolation: Principles and Practice', G.D. Foster & D. Twell editors, John Wiley & Sons Ltd., pp.331-368。
Claims (17)
- 形質転換植物における遺伝子調節の特異性を改良する方法であって、
植物において、内在性ターゲット遺伝子の発現部位の一つにおける発現を含む、少なくとも2つの発現部位における遺伝子発現を生じさせるプロモーターと、前記内在性ターゲット遺伝子またはその産物の発現を調節する核酸配列である作用物質とを含むキメラ遺伝子を植物細胞に導入する工程、ならびに
前記植物細胞から形質転換植物を再生させる工程を含み、
ここで、前記形質転換植物は前記キメラ遺伝子を含み、前記形質転換植物における前記キメラ遺伝子の発現により、前記キメラ遺伝子と前記内在性ターゲット遺伝子とがオーバーラップする発現部位で、前記内在性ターゲット遺伝子またはその産物が調節されるものであるとともに、
前記キメラ遺伝子と前記内在性ターゲット遺伝子との共同発現の部位が、前記キメラ遺伝子及び前記内在性ターゲット遺伝子の各々が単独で発現する部位よりも、より限定されている、前記方法。 - 前記核酸配列がコードまたは非コード配列である、請求項1に記載の方法。
- 前記ターゲット遺伝子の発現がアップレギュレートされる、請求項1または2に記載の方法。
- 前記ターゲット遺伝子の発現がダウンレギュレートされる、請求項1または2に記載の方法。
- 前記核酸配列がアンチセンス方向のものである、請求項1、2または4に記載の方法。
- 前記核酸がセンス方向のものである、請求項1〜4の何れか1項に記載の方法。
- 前記ターゲット遺伝子のアップレギュレーションが、前記ターゲット遺伝子のプロモーターのアクチベーターの導入から生じるものである、請求項3に記載の方法。
- 前記キメラ遺伝子の前記作用物質が複数の核酸配列を含み、その配列のそれぞれは、発現された場合に特定の機能を果たすものである、請求項1〜7のいずれか1項に記載の方法。
- 前記キメラ遺伝子のプロモーターが、前記内在性遺伝子の複数のオーバーラップ発現部位で発現される、請求項1〜8のいずれか1項に記載の方法。
- 前記プロモーターがKNT1プロモーターまたはRB7プロモーターである、請求項1〜9のいずれか1項に記載の方法。
- 前記核酸配列が、1以上のリボザイム、メッセンジャーRNAを分解するための標的RNアーゼ、または特異的RNAの安定化もしくは不安定化物質である、請求項1〜9のいずれか1項に記載の方法。
- 前記核酸が転写のアクチベーターを含む、請求項1〜9のいずれか1項に記載の方法。
- 転写の前記アクチベーターがタンパク質をコードする熱ショック因子である、請求項12に記載の方法。
- 前記作用物質が前記遺伝子上で作用する機構が、アンチセンス、共同抑制、ターゲット遺伝子のプロモーターの阻害もしくは活性化、転写の阻害もしくは活性化、メッセンジャーRNA安定性の変更もしくはmRNAの分解、翻訳の阻害もしくは活性化、タンパク質の阻害もしくは活性化、タンパク質代謝回転の変更、共同因子としての作用、タンパク質−タンパク質相互作用の変更、または生化学的経路によるフラックスの変更からなる群の1以上である、請求項1〜13のいずれか1項に記載の方法。
- 受託番号NCIMB 40871の寄託された構築物。
- プラスミドpATC37010またはそのプラスミドに含まれるようなプロモーター−遺伝子断片−ターミネーターカセットを含む構築物。
- 請求項1〜14のいずれか1項に記載の方法を行うことにより得られた植物およびその子孫。
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GBGB9706381.2A GB9706381D0 (en) | 1997-03-27 | 1997-03-27 | Improvements relating to the specificity of gene expression |
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JP51977498A Pending JP2001517076A (ja) | 1997-03-27 | 1998-03-27 | 遺伝子発現の特異性に関する改良 |
JP2009211993A Ceased JP2010029196A (ja) | 1997-03-27 | 2009-09-14 | 遺伝子発現の特異性に関する改良 |
JP2013232689A Pending JP2014057593A (ja) | 1997-03-27 | 2013-11-11 | 遺伝子発現の特異性に関する改良 |
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US (1) | US6140554A (ja) |
EP (1) | EP0977877B1 (ja) |
JP (3) | JP2001517076A (ja) |
CN (1) | CN1161474C (ja) |
AR (1) | AR010410A1 (ja) |
AT (1) | ATE327337T1 (ja) |
AU (1) | AU750951B2 (ja) |
BR (1) | BR9808430B1 (ja) |
CA (1) | CA2285221C (ja) |
DE (1) | DE69834637T2 (ja) |
DK (1) | DK0977877T3 (ja) |
ES (1) | ES2265660T3 (ja) |
GB (1) | GB9706381D0 (ja) |
HK (1) | HK1025792A1 (ja) |
NZ (1) | NZ337646A (ja) |
WO (1) | WO1998044138A1 (ja) |
ZA (1) | ZA982568B (ja) |
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AUPP249298A0 (en) | 1998-03-20 | 1998-04-23 | Ag-Gene Australia Limited | Synthetic genes and genetic constructs comprising same I |
CZ295108B6 (cs) | 1998-03-20 | 2005-05-18 | Benitec Australia Ltd | Syntetický gen obsahující dispergovanou nebo cizorodou deoxyribonukleovou molekulu a genový konstrukt obsahující tento syntetický gen |
EP2314700A1 (en) | 1999-01-28 | 2011-04-27 | Medical College of Georgia Research Institute, Inc | Composition and method for in vivo and in vitro attenuation of gene expression using double stranded RNA |
FR2794772B1 (fr) * | 1999-06-10 | 2001-09-21 | Agronomique Inst Nat Rech | Promoteur permettant l'expression de transgenes dans toute la plante hormis dans la graine |
US6423885B1 (en) | 1999-08-13 | 2002-07-23 | Commonwealth Scientific And Industrial Research Organization (Csiro) | Methods for obtaining modified phenotypes in plant cells |
WO2002029028A2 (en) * | 2000-10-03 | 2002-04-11 | Bayer Bioscience N.V. | Brassicaceae with altered floral development |
GB0025225D0 (en) * | 2000-10-14 | 2000-11-29 | Cambridge Advanced Tech | Plant cell death system |
US20050066394A1 (en) * | 2000-11-28 | 2005-03-24 | Olga Danilevskaya | Floral development genes |
HUP0501003A3 (en) * | 2000-11-28 | 2008-03-28 | Du Pont | Floral development genes |
CA2369944A1 (en) | 2001-01-31 | 2002-07-31 | Nucleonics Inc. | Use of post-transcriptional gene silencing for identifying nucleic acid sequences that modulate the function of a cell |
DE10325814A1 (de) * | 2003-06-06 | 2004-12-23 | Icon Genetics Ag | Sichere Erzeugung eines gewünschten Produkts in hybriden Samen |
US8304607B2 (en) * | 2004-04-20 | 2012-11-06 | Syngenta Participations Ag | Epidermal tissue promoter derived from potato for monocots |
US9238818B2 (en) * | 2004-04-20 | 2016-01-19 | Syngenta Participations Ag | Methods and genetic constructs for modification of lignin composition of corn cobs |
US8129588B2 (en) | 2004-04-20 | 2012-03-06 | Syngenta Participations Ag | Regulatory sequences for expressing gene products in plant reproductive tissue |
GB0421598D0 (en) | 2004-09-29 | 2004-10-27 | Cambridge Advanced Tech | Modification of plant development and morphology |
EP1874936B1 (en) | 2005-04-19 | 2013-10-30 | BASF Plant Science GmbH | Improved methods controlling gene expression |
WO2018089099A1 (en) | 2016-10-11 | 2018-05-17 | Dow Agrosciences Llc | Modulation of transgene expression in plants |
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1997
- 1997-03-27 GB GBGB9706381.2A patent/GB9706381D0/en active Pending
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1998
- 1998-03-26 ZA ZA9802568A patent/ZA982568B/xx unknown
- 1998-03-27 EP EP98913933A patent/EP0977877B1/en not_active Revoked
- 1998-03-27 CN CNB988054566A patent/CN1161474C/zh not_active Expired - Fee Related
- 1998-03-27 AU AU68453/98A patent/AU750951B2/en not_active Ceased
- 1998-03-27 WO PCT/GB1998/000939 patent/WO1998044138A1/en active IP Right Grant
- 1998-03-27 NZ NZ337646A patent/NZ337646A/en not_active IP Right Cessation
- 1998-03-27 CA CA2285221A patent/CA2285221C/en not_active Expired - Fee Related
- 1998-03-27 BR BRPI9808430-5A patent/BR9808430B1/pt not_active IP Right Cessation
- 1998-03-27 DE DE69834637T patent/DE69834637T2/de not_active Revoked
- 1998-03-27 US US09/049,475 patent/US6140554A/en not_active Expired - Lifetime
- 1998-03-27 AT AT98913933T patent/ATE327337T1/de not_active IP Right Cessation
- 1998-03-27 ES ES98913933T patent/ES2265660T3/es not_active Expired - Lifetime
- 1998-03-27 AR ARP980101420A patent/AR010410A1/es active IP Right Grant
- 1998-03-27 JP JP51977498A patent/JP2001517076A/ja active Pending
- 1998-03-27 DK DK98913933T patent/DK0977877T3/da active
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- 2009-09-14 JP JP2009211993A patent/JP2010029196A/ja not_active Ceased
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- 2013-11-11 JP JP2013232689A patent/JP2014057593A/ja active Pending
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Also Published As
Publication number | Publication date |
---|---|
BR9808430B1 (pt) | 2010-10-19 |
JP2001517076A (ja) | 2001-10-02 |
CA2285221A1 (en) | 1998-10-08 |
ATE327337T1 (de) | 2006-06-15 |
CA2285221C (en) | 2010-10-12 |
AR010410A1 (es) | 2000-06-07 |
ZA982568B (en) | 2000-10-23 |
WO1998044138A1 (en) | 1998-10-08 |
GB9706381D0 (en) | 1997-05-14 |
HK1025792A1 (en) | 2000-11-24 |
DE69834637D1 (de) | 2006-06-29 |
AU750951B2 (en) | 2002-08-01 |
EP0977877A1 (en) | 2000-02-09 |
EP0977877B1 (en) | 2006-05-24 |
AU6845398A (en) | 1998-10-22 |
ES2265660T3 (es) | 2007-02-16 |
BR9808430A (pt) | 2000-05-23 |
DK0977877T3 (da) | 2006-09-25 |
US6140554A (en) | 2000-10-31 |
NZ337646A (en) | 2001-06-29 |
CN1161474C (zh) | 2004-08-11 |
CN1257548A (zh) | 2000-06-21 |
JP2014057593A (ja) | 2014-04-03 |
DE69834637T2 (de) | 2006-10-05 |
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