JP2015509737A - 改善された糖化特性を有する遺伝子組換え植物 - Google Patents
改善された糖化特性を有する遺伝子組換え植物 Download PDFInfo
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- C12N15/8245—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving modified carbohydrate or sugar alcohol metabolism, e.g. starch biosynthesis
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- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
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- C12N15/8255—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving lignin biosynthesis
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- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
- C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
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- C12N15/8282—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance for fungal resistance
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- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/18—Carboxylic ester hydrolases (3.1.1)
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- C12Y301/00—Hydrolases acting on ester bonds (3.1)
- C12Y301/01—Carboxylic ester hydrolases (3.1.1)
- C12Y301/01072—Acetylxylan esterase (3.1.1.72)
Abstract
Description
(a)配列番号1のヌクレオチド配列を含むポリヌクレオチド、
(b)ストリンジェントなハイブリダイゼーション条件下で、(a)に定義されているポリヌクレオチドのポリペプチドコード領域に相補的なヌクレオチド配列にハイブリダイズすることができ、生物活性を有するアセチルキシランエステラーゼポリペプチドまたはその機能的に均等な改変型をコードするヌクレオチド配列を含むポリヌクレオチド、および
(c)(a)または(b)に定義されているヌクレオチド配列に対する遺伝暗号の結果としての縮重であり、生物活性を有するアセチルキシランエステラーゼポリペプチドまたはその機能的に均等な改変型をコードする核酸配列を含むポリヌクレオチド
から選択される。
(a)配列番号2、3、4または5として示すアミノ酸配列を含むポリペプチド、
(b)配列番号2、3、4または5として示すアミノ酸配列から本質的になるポリペプチド、および
(c)配列番号2として示すアミノ酸配列からなるポリペプチド
から選択される。
(実施例1)
アセチルキシランエステラーゼ遺伝子による植物の形質転換
次のプライマーを用いて、クロコウジカビアセチルキシランエステラーゼをコードするcDNA(配列番号1)を増幅した。
5'Fc2fuf(caccATGCTATCAACCCACCTCCTCTCGC)(配列番号6)および
3'Fc2rls(TCAAGCAAACCCAAACCACTCCATATCCTTATC)(配列番号7)
シロイヌナズナにおけるCE1発現の効果
シロイヌナズナにおいて、4種の独立的な、単一挿入のホモ接合体系統を解析した。逆転写ポリメラーゼ連鎖反応(RT-PCR)により、導入遺伝子の発現を検出した。系統6cは、最高の転写物レベルを有していた(図1)。
雑種アスペンにおけるCE1発現の効果
雑種アスペン(ヨーロッパヤマナラシ(Populus tremula)×アメリカヤマナラシ(tremuloides))、クローンT89において、同一真菌構築物による同様の実験を行った。キシランエステラーゼ転写物が存在するトランスジェニック系統を得て(図13)、繁殖させ、温室内に植えた。本出願人らが、基質として基質p-ナフチルアセテートを用いてトランスジェニック系統におけるエステラーゼ活性を検査したとき、全系統は、WTと比較して酵素活性の増加を示し、活性の増加は、壁結合画分に見られた(図14)。一部の系統において、植物成長は少々影響されていた(図15)。高さ成長は、酵素活性とは全く関係なく、系統5、8および11で増加し、系統4で減少した。直径成長は、有意に影響されなかった。
Claims (16)
- 植物における糖化能力を増加させるための方法であって、前記植物における少なくとも1種の細胞型において、アセチルキシランエステラーゼポリペプチドをコードするポリヌクレオチドを過剰発現させる工程を含む方法。
- 前記植物におけるグルコース収量を改善するための、請求項1に記載の方法。
- 遺伝子組換え植物を作出するための方法であって、前記植物における少なくとも1種の細胞型において、アセチルキシランエステラーゼポリペプチドをコードするポリヌクレオチドを過剰発現させる工程を含む方法。
- 前記植物が、対応する非遺伝子組換え野生型植物と比較して増加した糖化能力を有する、請求項3に記載の方法。
- 植物細胞において機能的なプロモーターであって、アセチルキシランエステラーゼポリペプチドをコードするポリヌクレオチドに作動可能に連結し、過剰発現を調節するプロモーターを含む発現カセットにより、前記細胞型を形質転換する工程を含む、請求項1から4のいずれか一項に記載の方法。
- 前記プロモーターが、CaMV 35Sプロモーターである、請求項5に記載の方法。
- 前記ポリヌクレオチドが、
(a)配列番号1のヌクレオチド配列を含むポリヌクレオチド、
(b)ストリンジェントなハイブリダイゼーション条件下で、(a)に定義されているポリヌクレオチドのポリペプチドコード領域に相補的なヌクレオチド配列にハイブリダイズすることができ、生物活性を有するアセチルキシランエステラーゼポリペプチドまたはその機能的に均等な改変型をコードするヌクレオチド配列を含むポリヌクレオチド、および
(c)(a)または(b)に定義されているヌクレオチド配列に対する遺伝暗号の結果としての縮重であり、生物活性を有するアセチルキシランエステラーゼポリペプチドまたはその機能的に均等な改変型をコードする核酸配列を含むポリヌクレオチド
から選択される、請求項1から6のいずれか一項に記載の方法。 - 前記アセチルキシランエステラーゼポリペプチドが、
(a)配列番号2、3、4または5として示すアミノ酸配列を含むポリペプチド、
(b)配列番号2、3、4または5として示すアミノ酸配列から本質的になるポリペプチド、および
(c)配列番号2、3、4または5として示すアミノ酸配列からなるポリペプチド
から選択される、請求項1から7のいずれか一項に記載の方法。 - 植物が、その細胞壁にアセチル化キシランを有する被子植物および他の植物から選択される、請求項1から8のいずれか一項に記載の方法。
- 植物が、ヤナギ科(family Salicaceae)のものである、請求項9に記載の方法。
- 植物または植物の部分が、酵素による加水分解の前に、酸またはアルカリ等の適した薬剤で前処理される、請求項1から10のいずれか一項に記載の方法。
- 前記植物における病原体に対する抵抗性を増加させるための、請求項1から11のいずれか一項に記載の方法。
- 請求項3から12のいずれか一項に記載の方法により作出された遺伝子組換え植物。
- 前記植物における少なくとも1種の細胞型において、アセチルキシランエステラーゼポリペプチドをコードするポリヌクレオチドを過剰発現する遺伝子組換え植物。
- 対応する非遺伝子組換え野生型植物と比較して増加した糖化能力を有する、請求項14に記載の遺伝子組換え植物。
- 対応する非遺伝子組換え野生型植物と比較して増加したグルコース収量を有する、請求項13から15のいずれか一項に記載の遺伝子組換え植物。
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
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US201261610963P | 2012-03-14 | 2012-03-14 | |
SE1250240 | 2012-03-14 | ||
SE1250240-7 | 2012-03-14 | ||
US61/610,963 | 2012-03-14 | ||
PCT/SE2013/050226 WO2013137810A1 (en) | 2012-03-14 | 2013-03-12 | Genetically modified plants having improved saccharification properties |
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JP2015509737A true JP2015509737A (ja) | 2015-04-02 |
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JP2015500393A Ceased JP2015509737A (ja) | 2012-03-14 | 2013-03-12 | 改善された糖化特性を有する遺伝子組換え植物 |
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Country | Link |
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US (1) | US20150033411A1 (ja) |
EP (1) | EP2825658A4 (ja) |
JP (1) | JP2015509737A (ja) |
CN (1) | CN104520430A (ja) |
AU (1) | AU2013232818A1 (ja) |
BR (1) | BR112014021981A2 (ja) |
CA (1) | CA2865805A1 (ja) |
CL (1) | CL2014002418A1 (ja) |
NZ (1) | NZ629407A (ja) |
WO (1) | WO2013137810A1 (ja) |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
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WO2009108941A2 (en) * | 2008-02-29 | 2009-09-03 | University Of Central Florida Research Foundation, Inc. | Production and use of plant degrading materials |
WO2009146464A2 (en) * | 2008-05-30 | 2009-12-03 | Edenspace Systems Corporation | Methods and compositions for modifying levels of lignin and increasing the digestibility of cellulose by the recombinant overexpression of lignocellulolytic enzymes |
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US8071335B2 (en) * | 2008-07-31 | 2011-12-06 | Novozymes, Inc. | Polypeptides having acetylxylan esterase activity and polynucleotides encoding same |
WO2012000888A1 (en) * | 2010-06-29 | 2012-01-05 | Dsm Ip Assets B.V. | Polypeptide having acetyl xylan esterase activity and uses thereof |
BR112013010825A2 (pt) * | 2010-11-03 | 2019-09-24 | Futuragene Israel Ltda | plantas transgênicas com rendimentos melhorados de sacarificação e métodos para geração das mesmas |
WO2012068310A2 (en) * | 2010-11-16 | 2012-05-24 | Qteros, Inc. | Compositions and methods for improved saccharification of genetically modified plant-derived biomass |
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- 2013-03-12 JP JP2015500393A patent/JP2015509737A/ja not_active Ceased
- 2013-03-12 EP EP13760377.5A patent/EP2825658A4/en not_active Withdrawn
- 2013-03-12 CA CA2865805A patent/CA2865805A1/en not_active Abandoned
- 2013-03-12 CN CN201380014014.2A patent/CN104520430A/zh active Pending
- 2013-03-12 AU AU2013232818A patent/AU2013232818A1/en not_active Abandoned
- 2013-03-12 BR BR112014021981A patent/BR112014021981A2/pt not_active Application Discontinuation
- 2013-03-12 NZ NZ629407A patent/NZ629407A/en not_active IP Right Cessation
- 2013-03-12 US US14/384,649 patent/US20150033411A1/en not_active Abandoned
- 2013-03-12 WO PCT/SE2013/050226 patent/WO2013137810A1/en active Application Filing
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2014
- 2014-09-12 CL CL2014002418A patent/CL2014002418A1/es unknown
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Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009108941A2 (en) * | 2008-02-29 | 2009-09-03 | University Of Central Florida Research Foundation, Inc. | Production and use of plant degrading materials |
WO2009146464A2 (en) * | 2008-05-30 | 2009-12-03 | Edenspace Systems Corporation | Methods and compositions for modifying levels of lignin and increasing the digestibility of cellulose by the recombinant overexpression of lignocellulolytic enzymes |
Non-Patent Citations (5)
Title |
---|
DATABASE UNIPROT [ONLINE], ACCESSIN NO. G3XVM1, <URL: HTTP://WWW.UNIPROT.ORG/UNIPROT/G3XVM1.TXT?VERS, JPN6016047659, ISSN: 0003458701 * |
DATABASE UNIPROT [ONLINE], ACCESSIN NO. G7XHV5, <URL: HTTP://WWW.UNIPROT.ORG/UNIPROT/G7XHV5.TXT?VERS, JPN6016047664, ISSN: 0003458703 * |
DATABASE UNIPROT [ONLINE], ACCESSIN NO. Q92194, <URL: HTTP://WWW.UNIPROT.ORG/UNIPROT/Q92194.TXT?VERS, JPN6016047666, ISSN: 0003458704 * |
DATABASE UNIPROT [ONLINE], ACCESSIN NO. Q96W96, <URL: HTTP://WWW.UNIPROT.ORG/UNIPROT/Q96W96.TXT?VERS, JPN6016047662, ISSN: 0003458702 * |
PLANT MOL. BIOL. (2011) VOL.77, ISSUE 4-5, P.433-445, JPN6016047657, ISSN: 0003458700 * |
Also Published As
Publication number | Publication date |
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CA2865805A1 (en) | 2013-09-19 |
AU2013232818A1 (en) | 2014-09-18 |
CN104520430A (zh) | 2015-04-15 |
EP2825658A1 (en) | 2015-01-21 |
CL2014002418A1 (es) | 2015-01-16 |
BR112014021981A2 (pt) | 2017-07-11 |
NZ629407A (en) | 2016-09-30 |
US20150033411A1 (en) | 2015-01-29 |
EP2825658A4 (en) | 2015-12-16 |
WO2013137810A1 (en) | 2013-09-19 |
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