JP5513883B2 - トランスジェニック事象mon89034に対応するトウモロコシ植物および種子ならびにそれらを検出および使用するための方法 - Google Patents
トランスジェニック事象mon89034に対応するトウモロコシ植物および種子ならびにそれらを検出および使用するための方法 Download PDFInfo
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- JP5513883B2 JP5513883B2 JP2009513394A JP2009513394A JP5513883B2 JP 5513883 B2 JP5513883 B2 JP 5513883B2 JP 2009513394 A JP2009513394 A JP 2009513394A JP 2009513394 A JP2009513394 A JP 2009513394A JP 5513883 B2 JP5513883 B2 JP 5513883B2
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Description
本願は、2006年5月26日に出願された米国仮出願番号60/808,834に対して優先権の利益を主張するものである。
本発明は、トランスジェニックトウモロコシ事象(event)MON89034ならびにその植物体の一部および種子に関する。この事象は、鱗翅(Lepidoptera)目の昆虫による昆虫侵入に抵抗性を示す。本発明はまた、このトランスジェニック事象に特有のヌクレオチド配列の存在について探索するときにこのトランスジェニック事象の存在についての診断であるDNAを含む植物および種子を使用するための方法、ならびに、このトランスジェニック事象に特有の特異的なヌクレオチド配列を検出することによって生物学的サンプル中の前記トウモロコシ事象の存在を検出するための方法にも関する。本発明は、この事象に特有のヌクレオチド配列を提供する。
本発明は、本明細書中で事象MON89034と呼ばれるトウモロコシ(Zea mays)植物の鱗翅目(Lepidopteran)抵抗性トランスジェニック品種に関し、また、トウモロコシの任意のサンプルまたは品種において検出されるときに、そのサンプルまたは品種中に存在する、トランスジェニックトウモロコシ植物事象MON89034の存在についての診断である特有のDNA配列に関し、そして、トウモロコシMON89034ならびにそれから得られる子孫植物および種子における導入遺伝子/ゲノム挿入領域の検出にも関する。
特許文献2:米国特許第6,620,988号明細書
非特許文献2:Roush(1994)Biocontrol. Sci. Technol.. 4:501-516
非特許文献3:Weising et al. (19880 Ann. Rev. Genet 22:421-477
本発明は、American Type Culture Collection(ATCC)に受入番号PTA-7455として2006年3月28日に寄託された種子を有する、MON89034と命名されたトランスジェニックトウモロコシ植物およびその(挿入されたトランスジェニックDNAに相当する少なくとも1つの対立遺伝子も含む限り)トウモロコシ事象MON89034と識別不能な子孫に関する。本発明の別の態様は、配列番号:1、配列番号:2、配列番号:3、配列番号:4および配列番号:5よりなる群から選択されるポリヌクレオチドを含む、子孫植物もしくは種子またはトウモロコシ事象MON89034の植物および種子の再生可能な一部である。本発明はまた、トウモロコシ事象MON89034の植物体の一部分を含み、その植物体の一部分が、上で示したようなポリヌクレオチドを少なくとも含む限り、これらの一部分として、花粉、胚珠、花、苗条、根、柄、花柱、雄穂、穂および葉が挙げられるが、これらに限定されない。MON89034事象に対応するトウモロコシ植物の分野におけるMON89034のゲノム内に含まれる新規遺伝的組成物およびMON89034からの製品(例えば、粗挽き粉、小麦粉、油、パルプおよび残渣バイオマス(biomass left over))は、本発明の一態様である。
以下の定義および方法は、本発明をよりよく定義するため、および、本発明を実施する際に当業者を導くために提供される。別段述べられない限り、用語は、関連分野の当業者による慣例的用法に従って理解されるべきである。分子生物学における一般的な用語の定義は、Rieger et al., Glossary of Genetics:Classical and Molecular,5th edition, Springer-Verlag:New York,1991;およびLewin, Genes V, Oxford University Press:New York,1994内に見られることがある。
(a)配列番号:5に示されるようなヌクレオチド配列;
(b)配列番号:1および配列番号:2に示されるようなヌクレオチド配列の両方;
(c)配列番号:3に示されるようなヌクレオチド配列;および
(d)配列番号:4に示されるようなヌクレオチド配列
よりなる群から選択される配列のうちのいずれか1つまたはすべてを含む。
この実施例では、トランスジェニックトウモロコシ事象MON89034の構築および分子の特徴づけを説明する。
この実施例では、ゲノム内のトランスジェニックトウモロコシ事象MON89034についての診断であるDNAを含むトウモロコシ植物の同定およびそのようなトウモロコシ植物の接合状態の判定を説明する。
Claims (23)
- 配列番号:2に示されるヌクレオチド配列またはその相補体、ならびに、Cry1A.105コード配列およびCry2Ab2コード配列を含むDNA分子。
- 配列番号:4に示されるヌクレオチド配列またはその相補体を含む、請求項1のDNA分子。
- 配列番号:1に示されるヌクレオチド配列またはその相補体をさらに含む、請求項1のDNA分子。
- 配列番号:3に示されるヌクレオチド配列またはその相補体を含む、請求項3のDNA分子。
- 配列番号:5で示されるヌクレオチド配列またはその相補体を含む、請求項1から4のいずれかに記載のDNA分子。
- 請求項1から5のいずれかに記載のDNA分子を含むトランスジェニックトウモロコシ植物細胞。
- 請求項1から5のいずれかに記載のDNA分子を含むトランスジェニックトウモロコシ植物またはその一部。
- 請求項7に記載のトランスジェニックトウモロコシ植物の種子であって、当該DNA分子を含む種子。
- トランスジェニックトウモロコシ植物またはその一部であって、前記トウモロコシ植物の種子は、受入番号PTA-7455としてAmerican Type Culture Collectionに寄託されている、トランスジェニックトウモロコシ植物またはその一部。
- Cry2AbをコードするDNAならびに配列番号:1および配列番号:2のヌクレオチド配列を有するDNAが、前記トウモロコシ植物またはその一部の細胞におけるゲノムの一部を形成している、昆虫抵抗性トウモロコシ植物またはその一部。
- Cry1A.105をコードするDNAをさらに含む、請求項10に記載の昆虫抵抗性トウモロコシ植物またはその一部。
- 請求項10または11に記載の昆虫抵抗性トウモロコシ植物の種子であって、Cry2AbをコードするDNAならびに配列番号:1および配列番号:2のヌクレオチド配列を有するDNAを含む、種子。
- 請求項7に記載のトランスジェニックトウモロコシ植物またはその一部に由来する組成物であって、検出可能な量の前記DNA分子を含み、コーンミール、コーン油、コーンケーキ、トウモロコシ種子、トウモロコシ胚芽、コーンスターチ、トウモロコシ粉、トウモロコシ花粉、トウモロコシ花柱、コーン浸出液、コーンモルト、コーンシュガー、コーンシロップ、コーン油から生産されるマーガリン、乾燥蒸留粕(DDGS)、化粧品およびバルキング剤よりなる群から選択される商品生産物である、組成物。
- 昆虫抵抗性トウモロコシ植物を作出する方法であって、
(a)請求項7に記載のトランスジェニックトウモロコシ植物と、異なるトウモロコシ植物とを交配し;
(b)工程(a)の交配に由来する少なくともひとつの子孫植物を得;ついで、
(c)配列番号:1および配列番号:2に示されるヌクレオチド配列を含む子孫を選択することを含み、ここに、選択される子孫が昆虫抵抗性トウモロコシ植物である、方法。 - 昆虫抵抗性トウモロコシ植物を作出する方法であって、
(a)トウモロコシ植物細胞を請求項1から5のいずれかに記載のDNA分子で形質転換し;ついで、
(b)前記DNA分子を含み、かつ、昆虫抵抗性であるトウモロコシ植物を、前記形質転換細胞から再生することを含む方法。 - 昆虫の侵入からトウモロコシ植物を保護するための方法であって、鱗翅目害虫の食餌中に、請求項7、9、10および11のいずれかに記載のトランスジェニックトウモロコシ植物またはその一部の細胞または組織を殺虫的に有効な量提供することを特徴とする方法。
- 前記鱗翅目害虫が、ヨトウムシ(Spodoptera frugiperda)、アワノメイガ(Ostrinia nubilalis)、オオタバコガ(Helicoverpa zea)、トウモロコシノメイガ(Diatraea grandiosella)およびタマナヤガ(Agrotis ipsilon)よりなる群から選択される請求項16に記載の方法。
- 生物学的サンプル中の配列番号:3、配列番号:4および配列番号:5よりなる群から選択されるDNA分子の存在を検出する方法であって、
(a)生物学的サンプルを、配列番号:3もしくはその相補体、配列番号:4もしくはその相補体または配列番号:5もしくはその相補体の連続的な少なくとも15ヌクレオチドを含むヌクレオチドであって、トウモロコシ植物MON89034またはその子孫から抽出されたDNAに対する診断であるDNAプライマーまたはプローブとして機能する、DNAプライマー対と接触させ;
(b)核酸増幅反応条件に供し;
(c)核酸増幅反応を行って、DNAアンプリコン分子を生成し;ついで、
(d)前記DNAアンプリコン分子を検出することを含み、ここに、配列番号:1、配列番号:2およびそれらの相補体の少なくとも一つを含むアンプリコンの検出が、前記生物学的サンプル中の前記DNA分子の存在を示唆するものである方法。 - 前記生物学的サンプルがトウモロコシ植物から抽出されたDNAサンプルである、請求項18に記載の方法。
- 生物学的サンプルにおける配列番号:3、配列番号:4および配列番号:5よりなる群から選択されるDNAの存在を検出するためのキットであって、トウモロコシ事象MON89034および/またはその子孫に対して特異的なDNAプライマーまたはプローブとして機能するヌクレオチドであって、配列番号:3もしくはその相補体、配列番号:4もしくはその相補体または配列番号:5もしくはその相補体の連続的な少なくとも15ヌクレオチドを含む、少なくともひとつのDNA分子を含む、キット。
- 前記少なくともひとつのDNA分子が、配列番号:1、配列番号:2またはその相補体を含む、請求項20に記載のキット。
- 生物学的サンプル中のトウモロコシ事象MON89034を含むトウモロコシ植物のDNAの接合状態を決定する方法であって、
(a)前記サンプルを、(1)トウモロコシ事象MON89034 DNAを含む核酸増幅反応に用いるとき、トウモロコシ事象MON89034に対する診断である第1のアンプリコンを生成し、(2)トウモロコシ事象MON89034 DNA以外のトウモロコシゲノムDNAを含む核酸増幅反応に用いるとき、トウモロコシ事象MON89034 DNA以外のトウモロコシゲノムDNAに対する診断である第2のアンプリコンを生成する、配列番号:6、配列番号:7および配列番号:10のヌクレオチド配列を有するDNAを含むプライマーセットと接触させ;
(b)核酸増幅反応を行い;ついで、
(c)そのようにして生成されたアンプリコンを検出することを含み、ここに、両方のアンプリコンの存在の検出は、前記サンプルがトウモロコシ事象MON89034 DNAに対してヘテロ接合性であることを示唆するものであって、第1のアンプリコンのみの検出は、前記サンプルがトウモロコシ事象MON89034 DNAに対してホモ接合性であることを示唆するものである、方法。 - 前記プライマーセットが、さらに配列番号:14および配列番号:15のヌクレオチド配列を有するDNAを一緒に使用する、請求項22に記載の方法。
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