JP2011087581A - Aequoreacoerulescens由来の新規な蛍光タンパク質およびその使用方法 - Google Patents
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Abstract
【解決手段】Aequorea coerulescensから得られる新規な無色のGFP様タンパク質(acGFP)、ならびにその蛍光性および非蛍光性の変異体および誘導体をコードする核酸組成物、そしてまた、これらの核酸組成物によってコードされるペプチドおよびタンパク質。上記タンパク質、あるいは上記核酸組成物によってコードされるタンパク質は有色であり、かつ/または蛍光性であり、かつ/または光活性化が可能であり、従って、様々な異なる生物学的適用における、特に標識化のための上記タンパク質の使用。および、そのような生物学的適用において使用される上記タンパク質を含むキット。
【選択図】図1
Description
後、1回以上、一般的には少なくとも2回、の更なる追加免疫投与が行われる。免疫後、宿主からの血液が採取され、この場合、血清が血液細胞から分離される。得られた抗血清に存在する免疫グロブリンを、知られている方法を使用して、例えば、アンモニウム塩分画化、DEAEクロマトグラフィーなどを使用して精製することができる。
大きいhydromedusaの数個の試料が、2001年8月に、ウラジオストック近くの日本海のロシア沿岸で集められた。1組の特徴により、これらのクラゲをAequorea coerulescensとして同定することが可能である(Kramp、Dana.Rept.、72:201〜202(1968);PogodinおよびYakovlev、Rus.J.Mar.Biol.、25:417〜419(1999))。A.coerulescensおよびA.victoria(A.forscalea、A.aequoreaの異名)は類似しているが、それらの特徴のいくつかは非常に異なる。最も明白な差は触手の数である。すなわち、A.victoriaは放射状導管1つあたり1つの触手を有するだけであり、一方、A.coerulescensは、隣接する放射状導管の各対の間に4つ〜6つの触手を有する。
Claims (32)
- Aequorea coerulescensのペプチドをコードし、
(a)配列番号2のアミノ酸配列を含むタンパク質をコードする核酸;
(b)配列番号1のヌクレオチド配列を含む核酸
から選択される、単離された核酸。 - (a)部位特異的変異誘発および/またはランダム変異誘発の少なくとも1つを使用して請求項1に記載される核酸から誘導される核酸;
(b)配列番号4、6、8、10、12、14、16、18、20、22または24から選択されるアミノ酸配列をコードする核酸;
(c)配列番号3、5、7、9、11、13、15、17、19、21または23から選択されるヌクレオチド配列を含む核酸;
(d)遺伝暗号の縮重性のために上記(a)の核酸とは異なる核酸;
からなる群から選択される、単離された核酸。 - 蛍光タンパク質をコードする、請求項2に記載の単離された核酸。
- (a)請求項2に記載の核酸;および
(b)細胞において核酸を発現させるために必要な調節エレメント
を含む、発現カセット。 - (a)請求項3に記載の核酸;および
(b)細胞において核酸を発現させるために必要な調節エレメント
を含む、発現カセット。 - 請求項4に記載の発現カセットを含む、細胞またはその子孫。
- 請求項5に記載の発現カセットを含む、細胞またはその子孫。
- 請求項2に記載の核酸によってコードされる、単離されたペプチド。
- 請求項8に記載のペプチドに対して特異的に結合する抗体。
- 請求項8に記載のペプチドを含む、融合ペプチド。
- 請求項8に記載のペプチドを発現することができる、遺伝子組換え生物。
- 生物学的分子を標識または検出するための方法であって、生物学的分子を請求項8に記載のペプチドに結合させることを含む、方法。
- 細胞または細胞オルガネラを標識または検出するための方法であって、請求項8に記載のペプチドを細胞において産生させることを含む、方法。
- 遺伝子発現を検出するための方法であって、請求項8に記載のペプチドを細胞において産生させることを含む、方法。
- 請求項3に記載の核酸によってコードされる、単離されたペプチド。
- 請求項15に記載のペプチドに対して特異的に結合する抗体。
- 請求項15に記載のペプチドを組み込んだ、融合ペプチド。
- 請求項15に記載のペプチドを発現することができる、遺伝子組換え生物。
- 生物学的分子を標識または検出するための方法であって、生物学的分子を請求項15に記載のペプチドに結合させることを含む、方法。
- 細胞または細胞オルガネラを標識または検出するための方法であって、請求項15に記載のペプチドを細胞において産生させることを含む、方法。
- 遺伝子発現を検出するための方法であって、請求項15に記載のペプチドを細胞において産生させることを含む、方法。
- (a)配列番号2、4、6、8、10、12、14、16、18、20、22または24に由来する少なくとも100アミノ酸残基の長さのペプチドをコードする断片;あるいは
(b)配列番号1、3、5、7、9、11、13、15、17、19、21または23に由来する少なくとも300残基の長さのヌクレオチド配列と実質的に類似または同一である断片
からなる群から選択される、核酸断片。 - 請求項22に記載の核酸断片を含み、蛍光タンパク質をコードする、核酸分子。
- (a)請求項23に記載の核酸分子;および
(b)細胞において核酸断片を発現させるために必要な調節エレメント
を含む、発現カセット。 - 請求項24に記載の発現カセットを含む、細胞またはその子孫。
- 請求項23に記載の核酸断片によってコードされる、単離されたペプチド。
- 請求項26に記載のペプチドに対して特異的に結合する抗体。
- 請求項26に記載のペプチドを組み込んだ、融合ペプチド。
- 請求項26に記載のペプチドを発現することができる、遺伝子組換え生物。
- 生物学的分子を標識または検出するための方法であって、生物学的分子を請求項26に記載のペプチドに結合させることを含む、方法。
- 細胞または細胞オルガネラを標識または検出するための方法であって、請求項26に記載のペプチドを細胞において産生させることを含む、方法。
- 遺伝子発現を検出するための方法であって、請求項26に記載のペプチドを細胞において産生させることを含む、方法。
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ATE458816T1 (de) * | 2002-01-22 | 2010-03-15 | Evrogen Ip | Fluoreszierendes protein aus aequorea coerulescens und ihre verwendungen |
WO2004058973A1 (en) | 2002-12-26 | 2004-07-15 | Zakrytoe Aktsionernoe Obschestvo 'evrogen' | Fluorescent proteins from copepoda species and methods for using same |
WO2006060739A2 (en) | 2004-12-01 | 2006-06-08 | Proteologics, Inc. | Ubiquitin ligase assays and related reagents |
UA93922C2 (ru) * | 2009-03-20 | 2011-03-25 | Михаил Викторович Разуменко | Способ получения пептидов, которые специфически распознают клетки определенного типа, и предназначены для терапевтических целей |
US8435961B2 (en) | 2009-06-26 | 2013-05-07 | Massachusetts Institute Of Technology | Methods and compositions for increasing the activity of inhibitory RNA |
WO2010151664A2 (en) | 2009-06-26 | 2010-12-29 | Massachusetts Institute Of Technology | Compositions and methods for treating cancer and modulating stress granule formation |
US8268550B2 (en) * | 2009-06-26 | 2012-09-18 | Massachusetts Institute Of Technology | Compositions and methods for identification of PARP function, inhibitors, and activators |
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WO2011135040A1 (en) | 2010-04-30 | 2011-11-03 | F. Hoffmann-La Roche Ag | Fluorescent antibody fusion protein, its production and use |
US9134301B2 (en) * | 2010-11-22 | 2015-09-15 | Bio-Rad Laboratories, Inc. | Sorting of adherent cells by selective transformation of labels |
US20150168416A1 (en) | 2012-06-06 | 2015-06-18 | The United States Of America, As Represented By The Secretary, Dept. Of Health And Human Services | KITS FOR DETECTING AND MONITORING ENDOCRINE DISRUPTING CHEMICALS (EDCs) |
US20140024066A1 (en) * | 2012-06-20 | 2014-01-23 | California Institute Of Technology | Animal model having photo-activatable mitochondria |
RU2602452C2 (ru) * | 2014-05-14 | 2016-11-20 | Федеральное государственное бюджетное учреждение науки Институт биологии гена Российской академии наук | Способ выявления веществ и их композиций с противоопухолевой активностью |
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JPH10509881A (ja) * | 1994-11-10 | 1998-09-29 | ザ リージェンツ オブ ザ ユニバーシティー オブ カリフォルニア | 改変緑色蛍光タンパク質 |
JP2000503536A (ja) * | 1996-01-18 | 2000-03-28 | ユニバーシティ オブ フロリダ リサーチ ファウンデーション,インコーポレイテッド | ヒト化グリーン蛍光タンパク質遺伝子および方法 |
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EP1485481A2 (en) | 2004-12-15 |
US20060167225A1 (en) | 2006-07-27 |
CA2474108A1 (en) | 2003-07-31 |
US7897726B2 (en) | 2011-03-01 |
US20090148940A1 (en) | 2009-06-11 |
JP2005526495A (ja) | 2005-09-08 |
US7667016B2 (en) | 2010-02-23 |
ES2340154T3 (es) | 2010-05-31 |
WO2003062270B1 (en) | 2004-04-01 |
US7879988B2 (en) | 2011-02-01 |
IL162568A (en) | 2010-12-30 |
CA2474108C (en) | 2012-06-19 |
IL162568A0 (en) | 2005-11-20 |
DK1485481T3 (da) | 2010-06-07 |
US7432053B2 (en) | 2008-10-07 |
RU2004125585A (ru) | 2005-04-20 |
WO2003062270A8 (en) | 2004-11-04 |
WO2003062270A2 (en) | 2003-07-31 |
JP4510464B2 (ja) | 2010-07-21 |
RU2330886C2 (ru) | 2008-08-10 |
EP1485481B1 (en) | 2010-02-24 |
US20090148939A1 (en) | 2009-06-11 |
ATE458816T1 (de) | 2010-03-15 |
WO2003062270A3 (en) | 2003-11-27 |
DE60331422D1 (de) | 2010-04-08 |
AU2003208520B2 (en) | 2008-06-05 |
US20090137033A1 (en) | 2009-05-28 |
JP5465649B2 (ja) | 2014-04-09 |
JP2010046084A (ja) | 2010-03-04 |
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