JP6788582B2 - 植物におけるタンパク質生産の改変 - Google Patents
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H3/00—Processes for modifying phenotypes, e.g. symbiosis with bacteria
- A01H3/02—Processes for modifying phenotypes, e.g. symbiosis with bacteria by controlling duration, wavelength, intensity, or periodicity of illumination
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H3/00—Processes for modifying phenotypes, e.g. symbiosis with bacteria
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H3/00—Processes for modifying phenotypes, e.g. symbiosis with bacteria
- A01H3/04—Processes for modifying phenotypes, e.g. symbiosis with bacteria by treatment with chemicals
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- 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
- C12N15/8242—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
- C12N15/8243—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
- C12N15/8251—Amino acid content, e.g. synthetic storage proteins, altering amino acid biosynthesis
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- 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
- C12N15/8242—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
- C12N15/8257—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 for the production of primary gene products, e.g. pharmaceutical products, interferon
- C12N15/8258—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 for the production of primary gene products, e.g. pharmaceutical products, interferon for the production of oral vaccines (antigens) or immunoglobulins
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Description
a)植物または植物の一部を処理して、当該植物または当該植物の一部において二次葉(secondary leaf)生物体量(biomass)の生産を増加させること;
b)目的のタンパク質をコードするヌクレオチド配列であって、当該植物中で活性のある調節領域と動作可能に連結されたヌクレオチド配列を含む1つ以上の核酸を、当該植物または当該植物の一部に導入すること;
c)当該植物または当該植物の一部を、目的のタンパク質をコードするヌクレオチド配列の発現を可能にする条件下で培養し、それにより目的のタンパク質を生産すること、ここで、目的タンパク質の収率が、同じ条件で生育させたが二次葉生物体量を増やすための処理がされなかった同様の植物の同じ組織から取得された目的タンパク質の収率と比較して、増加している、
を含む方法を提供する。
本発明は、植物中でタンパク質を生産する方法に関する。本発明はまた、植物中で目的のタンパク質を生産するための方法および組成物を提供する。植物または植物の一部内で、目的のタンパク質を生産する方法もまた、提供される。
a)植物または植物の一部を処理して、当該植物または当該植物の一部において二次葉生物体量の生産を増加させること;
b)目的のタンパク質をコードするヌクレオチド配列であって、当該植物中で活性のある調節領域と動作可能に連結されたヌクレオチド配列を含む1つ以上の核酸を、当該植物または当該植物の一部に導入すること;
c)当該植物または当該植物の一部を、目的のタンパク質をコードするヌクレオチド配列の発現を可能にする条件下で培養し、それにより目的のタンパク質を生産すること、ここで、目的タンパク質の収率が、同じ条件で生育させたが二次生物体量を増やすための処理がされなかった同様の植物の同じ組織から取得される目的タンパク質の収率と比較して、増加している、
を含む方法を提供する。
a)植物または植物の一部を処理して、当該植物または当該植物の一部において二次葉生物体量の生産を増加させること;
b)ヘマグルチニン(HA)をコードするヌクレオチド配列であって、当該植物中で活性のある調節領域と動作可能に連結されたヌクレオチド配列を含む、1つ以上の核酸を、当該植物または当該植物の一部に導入すること;
c)当該植物または当該植物の一部を、HAをコードするヌクレオチド配列の発現を可能にする条件下で培養し、それによりHAを生産すること、ここで、HAの収率が、同じ条件で生育させたが二次生物体量を増やすための処理がされなかった同様の植物の同じ組織から取得されるHAの収率と比較して、増加している、
を含む方法が提供される。
(1)日中16時間/夜間8時間の光周期、80μmol/m2.sの光強度;
(2)日中16時間/夜間8時間の光周期、160μmol/m2.sの光強度;
(3)日中24時間の光周期、80μmol/m2.sの光強度;
(4)日中24時間の光周期、160μmol/m2.sの光強度。
(1)日中24時間の光周期、175μmol/m2.sの光強度(低光強度コントロール);
(2)日中24時間の光周期、350μmol/m2.sの光強度(高光強度試験)。
Claims (19)
- 植物または植物の一部内で目的の異種タンパク質を生産する方法であって:
a)植物または植物の一部を処理して、二次葉生物体量を増加させること、ここで、処理することは、植物の生長期間の明期を増加させること、植物の生長期間の光強度を増加させること、ホルモンの存在下で植物を培養すること、またはこれらの組み合わせを含み、処理された植物または植物の一部を生産する、ここで、明期および光強度の増加は、80μmol/m 2 .sの光強度で16/8光周期下で生長させた処理されていない植物と比較される;
b)目的の異種タンパク質をコードするヌクレオチド配列であって、当該植物中で活性のある調節領域と動作可能に連結されたヌクレオチド配列を含む1つ以上の核酸を、当該処理された植物または当該植物の一部に導入すること;および
c)当該処理された植物または当該植物の一部を、目的の異種タンパク質をコードするヌクレオチド配列の発現を可能にする条件下で培養し、それにより目的のタンパク質を生産すること、ここで、目的の異種タンパク質の収率が、当該処理されていない植物の植物の組織から取得される目的の異種タンパク質の収率と比較して、増加している、
を含み、
処理のステップである、ステップa)が、導入するステップである、ステップb)の約40日前から、導入するステップである、ステップb)の約9日前までに実施される、方法。 - 処理のステップである、ステップa)において、植物または植物の一部が、0.2:1および3:1の間の二次葉生物体量:一次葉生物体量の比を有する、請求項1に記載の方法。
- 植物または植物の一部を処理するステップである、ステップa)が、1つ以上の核酸を導入するステップの約20日前から、1つ以上の核酸を導入する日の約9日前までに実行される、請求項1に記載の方法。
- 植物が、導入するステップである、ステップb)の約9日前、約10日前、約11日前、約12日前、約13日前、または約14日前にホルモンの存在下で培養されるか、または、明期を増加させること、または光強度を増加させることが、導入するステップである、ステップb)の少なくとも約20日前に実施される、請求項1に記載の方法。
- さらに、植物または植物の一部を収穫するステップd)を含む、請求項1に記載の方法。
- さらに、植物または植物の一部を収穫して目的の異種タンパク質を精製するステップd)を含む、請求項1に記載の方法。
- 一次茎および二次茎が収穫される、請求項5に記載の方法。
- 二次茎が収穫される、請求項5に記載の方法。
- さらに、一次茎由来の中間の葉(P2)、および、一次茎由来の若葉(P1)が収穫される、請求項8に記載の方法。
- 一次茎由来の中間の葉(P2)、一次茎由来の若葉(P1)、二次茎由来の老葉(S3)、二次茎由来の中間の葉(S2)、および、二次茎由来の若葉(S1)が収穫される、請求項5に記載の方法。
- 当該植物の一次茎由来の老葉(P3)が、収穫から除外される、請求項5に記載の方法。
- ヌクレオチド配列が、薬剤的に活性のあるタンパク質、抗体、抗原、ワクチン、酵素、または工業用酵素をコードする、請求項1に記載の方法。
- ヌクレオチド配列が、インフルエンザHAタンパク質をコードする、請求項1に記載の方法。
- ステップc)において、核酸が植物または植物の一部中で一過性に発現される、請求項1に記載の方法。
- ステップc)において、核酸が植物または植物の一部中で安定的に発現される、請求項1に記載の方法。
- 植物から取得される目的の異種タンパク質が、HAタンパク質を含むウイルス様粒子(VLP)である、請求項13に記載の方法。
- 二次葉生物体量の増加が、二次茎および葉の数の増加、二次茎および葉の長さの増加、または両方の結果である、請求項1に記載の方法。
- ホルモンがサイトカイニンまたは合成サイトカイニンを含む、請求項1に記載の方法。
- 合成サイトカイニンが6−ベンジルアミノプリン(BAP)である、請求項18に記載の方法。
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