JPH11512441A - 緑色蛍光性タンパクであるgfpの新規な変種 - Google Patents
緑色蛍光性タンパクであるgfpの新規な変種Info
- Publication number
- JPH11512441A JPH11512441A JP9512326A JP51232697A JPH11512441A JP H11512441 A JPH11512441 A JP H11512441A JP 9512326 A JP9512326 A JP 9512326A JP 51232697 A JP51232697 A JP 51232697A JP H11512441 A JPH11512441 A JP H11512441A
- Authority
- JP
- Japan
- Prior art keywords
- gfp
- fluorescent protein
- cells
- fluorescent
- sequence
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
- C07K14/43595—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from coelenteratae, e.g. medusae
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/48—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
- Y10S530/808—Materials and products related to genetic engineering or hybrid or fused cell technology, e.g. hybridoma, monoclonal products
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
- Y10S530/855—Proteins from animals other than mammals or birds
- Y10S530/857—Fish; fish eggs; shell fish; crustacea
Abstract
Description
Claims (1)
- 【特許請求の範囲】 1.緑色蛍光性タンパク(GFP)またはその何れかの機能的類縁体から誘導 された蛍光性タンパクであって、蛍光強度を増大させるために、発色団に先行す る位置1にあるアミノ酸が突然変異されている蛍光性タンパク。 2.請求項1に記載の蛍光性タンパクであって、前記発色団がGFPの予想一 時アミノ酸配列の位置65−67にある蛍光性タンパク。 3.請求項1または2に記載の蛍光性タンパクであって、該蛍光性タンパクを 発現する細胞において、該細胞を 30℃以上の温度、好ましくは 32℃〜39℃の温 度、より好ましくは 35℃〜38℃の温度、最も好ましくは約 37℃の温度でインキ ュベートしたときに増大した蛍光を生じる蛍光性タンパク。 4.請求項1〜3の何れか1項に記載の蛍光性タンパクであって、該タンパク はアエクオレア・ビクトリア(Aequorea victorea)またはレニラ・レニホルミス (Renilla reniformis)由来である蛍光性タンパク。 5.請求項1〜4の何れか1項に記載の蛍光性タンパクであって、GFPまた は Y66H-GFP の位置64にあるアミノ酸Fが、L,I,V,AおよびGか らなる群から選択されるアミノ酸で置換されている蛍光性タンパク。 6.請求項1〜5の何れか1項に記載の蛍光性タンパクであって、前記発色団 に先行する位置1にあるアミノ酸FがLで置換されており、発色団のアミノ酸が SYG,SHGまたはTYGを含んでいる蛍光性タンパク。 7.請求項1〜6の何れか1項に記載の蛍光性タンパクであって、図3、図4 または図5のアミノ酸配列を有する蛍光性タンパク。 8.請求項1〜7の何れか1項に記載の蛍光性タンパク(GFP)を含む融合 化合物であって、前記GFPがポリペプチドに連結されている融合化合物。 9.請求項8に記載の融合化合物であって、前記ポリペプチドがキナーゼ、好 ましくはプロテインキナーゼA若しくはプロテインキナーゼCの触媒サブユニッ ト、Erkl、または細胞骨格要素である融合化合物。 10.請求項1,2,3,4,5,6若しくは7の蛍光性タンパクまたは請求 項7若しくは8の融合化合物をコードするヌクレオチド配列。 11.図3,図4および図5に示す配列から選択される、請求項10に記載の ヌクレオチド配列。 12.適切な1以上の制御領域と、請求項10または11のヌクレオチド配列 とを具備し、該配列は前記制御領域の制御下にあるDNA構築物 13.請求項12に記載のDNA構築物であって、天然のGFPプロモータ、 或いは哺乳類の構造的もしくは調節的プロモータ、ウイルスのプロモータ、酵母 のプロモータ、糸状菌のプロモータまたはバクテリアプロモータの制御下にある DNA構築物。 14.請求項12または13のDNA構築物で形質転換された宿主。 15.バクテリア、酵母、真菌、ブロトゾアおよび高度の真核生物から選択さ れる、請求項14に記載の宿主。 16.ポリペプチドを製造する方法であって、請求項14または15の宿主を 培養することと、該ホストから、前記ヌクレオチド配列によって発現されたポリ ペプチドを回収することとを具備した方法。 17.請求項16に記載の方法であって、前記ヌクレオチド配列の発現が、天 然のGFPプロモータによって行われる方法。 18.プロテインキナーゼ活性または脱リン酸化活性を測定するためのインビ トロ試験における、請求項1,2,3,4,5,6または7の蛍光性タンパクの 使用であって、純粋な形の前記蛍光性タンパクが生物学的サンプル、好ましくは 細胞抽出物に添加され、蛍光の何らかの変化が記録される使用。 19.代謝活性、好ましくはキナーゼ活性および脱リン酸化活性を測定するた めのインビボ試験における、請求項14または15の宿主の使用。 20.生きた細胞内における遺伝子発現のためのレポータとしての、請求項1 ,2,3,4,5,6または7の蛍光性タンパクの使用。 21.生きた完全な細胞内における2以上の遺伝子を同時にモニターするため の、請求項1,2,3,4,5,6または7の蛍光性タンパクの使用。 22.生きた細胞内における細胞小器官プロセスまたは細胞プロセスを同時に 可視化するための細胞小器官または細胞の標識としての、請求項1,2,3,4 ,5,6または7の蛍光性タンパクの使用。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DK1065/95 | 1995-09-22 | ||
DK106595 | 1995-09-22 | ||
PCT/DK1996/000051 WO1997011094A1 (en) | 1995-09-22 | 1996-01-31 | Novel variants of green fluorescent protein, gfp |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2003170625A Division JP2004024258A (ja) | 1995-09-22 | 2003-06-16 | 緑色蛍光性タンパクであるgfpの新規な変種 |
Publications (2)
Publication Number | Publication Date |
---|---|
JPH11512441A true JPH11512441A (ja) | 1999-10-26 |
JP3535177B2 JP3535177B2 (ja) | 2004-06-07 |
Family
ID=8100600
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP51232697A Expired - Lifetime JP3535177B2 (ja) | 1995-09-22 | 1996-01-31 | 緑色蛍光性タンパクであるgfpの新規な変種 |
JP2003170625A Pending JP2004024258A (ja) | 1995-09-22 | 2003-06-16 | 緑色蛍光性タンパクであるgfpの新規な変種 |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2003170625A Pending JP2004024258A (ja) | 1995-09-22 | 2003-06-16 | 緑色蛍光性タンパクであるgfpの新規な変種 |
Country Status (10)
Country | Link |
---|---|
US (3) | US6172188B1 (ja) |
EP (1) | EP0851874B1 (ja) |
JP (2) | JP3535177B2 (ja) |
AT (1) | ATE184613T1 (ja) |
AU (1) | AU4482996A (ja) |
CA (1) | CA2232727C (ja) |
DE (1) | DE69604298T2 (ja) |
DK (1) | DK0851874T3 (ja) |
ES (1) | ES2139329T3 (ja) |
WO (1) | WO1997011094A1 (ja) |
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WO1997011094A1 (en) | 1997-03-27 |
US6172188B1 (en) | 2001-01-09 |
DE69604298D1 (de) | 1999-10-21 |
ATE184613T1 (de) | 1999-10-15 |
US20050054050A1 (en) | 2005-03-10 |
DE69604298T2 (de) | 2000-05-18 |
CA2232727C (en) | 2002-03-26 |
US7314915B2 (en) | 2008-01-01 |
CA2232727A1 (en) | 1997-03-27 |
US6818443B2 (en) | 2004-11-16 |
EP0851874A1 (en) | 1998-07-08 |
JP3535177B2 (ja) | 2004-06-07 |
DK0851874T3 (da) | 2000-03-06 |
US20020107362A1 (en) | 2002-08-08 |
AU4482996A (en) | 1997-04-09 |
JP2004024258A (ja) | 2004-01-29 |
EP0851874B1 (en) | 1999-09-15 |
ES2139329T3 (es) | 2000-02-01 |
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