JP2001512474A - インビトロ・ペプチドまたはタンパク質発現ライブラリ - Google Patents
インビトロ・ペプチドまたはタンパク質発現ライブラリInfo
- Publication number
- JP2001512474A JP2001512474A JP53638498A JP53638498A JP2001512474A JP 2001512474 A JP2001512474 A JP 2001512474A JP 53638498 A JP53638498 A JP 53638498A JP 53638498 A JP53638498 A JP 53638498A JP 2001512474 A JP2001512474 A JP 2001512474A
- Authority
- JP
- Japan
- Prior art keywords
- protein
- dna
- library
- peptide
- binding
- 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
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
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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
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1034—Isolating an individual clone by screening libraries
- C12N15/1075—Isolating an individual clone by screening libraries by coupling phenotype to genotype, not provided for in other groups of this subclass
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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
- C12N2795/00—Bacteriophages
- C12N2795/00011—Details
- C12N2795/10011—Details dsDNA Bacteriophages
- C12N2795/10111—Myoviridae
- C12N2795/10122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
Abstract
Description
Claims (1)
- 【特許請求の範囲】 1. ペプチド又はタンパク質の多様な集団を表示し該ペプチド又はタンパク質 は、タンパク質:DNAの共役結合により特定のペプチド又はタンパク質のコード 化DNAと会合し少なくとも下記段階で構成されるペプチド又はタンパク質の発現 ライブラリを産生する方法。 1) アミノ酸配列をコード化するヌクレオチド配列を含み該アミノ酸配列は、 タンパク質:DNA共役結合(結合部分)、アミノ酸配列表示コード化配列 (表示部分)、及び、少なくとも一つの結合部分の付着サイトにより該コ ード化配列と結合する増幅可能なDNA分子の遺伝子ライブラリを調製する 。 2) このようにして形成された遺伝子ライブラリを発現する。 2. 一つの宿主細胞又は生物体当たり1つより多い任意のコピー数存在する単 一ライブラリ・メンバーにより、遺伝子物質をインビボで発現する請求項1に記 載の方法。 3. 特定コード化配列に結合するアミノ酸配列が、シス型として作用するタン パク質又は機能的にこれと同等のフラグメント、その変異体又はその誘導体から 誘導され、一つの宿主細胞又は生物体当たり1つより多い任意のコピー数存在す る少なくとも一つのライブラリ・メンバーにより、遺伝子物質の発現をインビボ で行う請求項1に記載の方法。 4. コード化配列に結合するアミノ酸配列がシス作用を行うタンパク質又はこ れと同等の機能を有するフラグメント、その変異体又はその誘導体から誘導され 、遺伝子物質の発現がインビトロで行われる請求項1に記載の方法。 5. シス作用タンパク質がP2Aタンパク質である請求項3又は4に記載の方法 。 6. 両側の付加サイトに隣接するDNAにはハイブリッド化するが付加サイトに 対応する領域ではハイブリッド化しないミスマッチ・オリゴヌクレオチドの存在 下で、当該発現を行わせる請求項4又は5に記載の方法。 7. 表示用のアミノ酸配列が最大40個までのアミノ酸残基である請求項1−6 のいずれか一つに記載の方法。 8. 表示用のアミノ酸配列をクローニングにより発生させ、又はクローニング に由来するDNAフラグメントで構成する請求項1−7のいずれか一つに記載の方 法。 9. 結合部分を、該結合部分が共有結合でコード化DNAに結合したまま残るよ うに修飾する請求項1−8のいずれか一つに記載の方法。 10.アミノ酸位置450にあるチロシンをフェニルアラニンで置換して修飾したP 2Aから、当該結合部分を誘導する請求項9に記載の方法。 11.請求項1−10のいずれか一つに記載の方法で産生した、インビトロ・ペプ チド発現ライブラリ。 12.アミノ酸配列をコード化する配列、タンパク質:DNAの共有結合(結合部 分)により当該コード化配列に特に結合するアミノ酸配列、アミノ酸配列を表示 するためにコード化する配列(表示部分)及び当該結合部分のための少なくとも 一つの付着サイト、並びに縮重配列及び/又は機能的に同等な配列を含み請求項 11に記載の、ペプチド又はタンパク質をライブラリ内で発現するためのコード化 DNA配列を含むDNA分子。 13.DNA配列を含む請求項12に記載のDNAベクター。 14.少なくとも下記の段階即ち、a)ライブラリを請求項11に記載のようにスク リーニングする段階、及び、b)該当するライブラリ・メンバーを選別し単離する 段階から成り、ペプチド又はタンパク質の発現ライブラリから、希望する性質を 示すライブラリ・メンバーを同定及び/又は精製する請求項11に記載の方法。 15.少なくとも下記の段階即ち、a)請求項11記載の方法で、標的分子を含むラ イブラリをスクリーニングする、及び、b)当該標的分子に結合しているライブラ リ・メンバーを選別し、単離する、及びに、c)特に当該標的分子に結合するペプ チド又はタンパク質を単離する。 から成る特定の標的結合ペプチド又はタンパク質を同定する方法。 16.特に標的分子に結合するペプチド又はタンパク質を追加的に発現するDNA を単離する請求項15に記載の方法。 17.a)サンプルに下記の(i)及び(ii)で構成される分子プローブで接触する。 (i)ペプチド又はタンパク質の標的結合部分で、請求項11記載の方法でライブラ リから選択したDNA部分、付加コード化DNAにより、標的分子に選択的に結合し得 るもの、及び(ii)リポーター部分。b)標的に結合したプローブを直接又は間接的 に評価する。 上記a)及びb)より成りサンプル中に標的分子が存在するかどうかを分析する方法 。 18.(i)請求項11に記載のライブラリから選択した付加コード化DNAで標的分子 に選択的に結合できるペプチド又はタンパク質部分(DNA部分)、及び(ii)リポ ータ部分より成り請求項17に記載の分析方法で使用する二官能性分子プローブ。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GB9703369.0 | 1997-02-18 | ||
GBGB9703369.0A GB9703369D0 (en) | 1997-02-18 | 1997-02-18 | Process |
PCT/GB1998/000518 WO1998037186A1 (en) | 1997-02-18 | 1998-02-18 | In vitro peptide or protein expression library |
Publications (2)
Publication Number | Publication Date |
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JP2001512474A true JP2001512474A (ja) | 2001-08-21 |
JP4644321B2 JP4644321B2 (ja) | 2011-03-02 |
Family
ID=10807887
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP53638498A Expired - Fee Related JP4644321B2 (ja) | 1997-02-18 | 1998-02-18 | インビトロ・ペプチドまたはタンパク質発現ライブラリ |
Country Status (14)
Country | Link |
---|---|
US (1) | US7416847B1 (ja) |
EP (1) | EP1007654B1 (ja) |
JP (1) | JP4644321B2 (ja) |
AT (1) | ATE297462T1 (ja) |
AU (1) | AU744791B2 (ja) |
CA (1) | CA2282497C (ja) |
DE (2) | DE1007654T1 (ja) |
DK (1) | DK1007654T3 (ja) |
ES (1) | ES2241117T3 (ja) |
GB (1) | GB9703369D0 (ja) |
IL (2) | IL131435A0 (ja) |
NO (1) | NO322653B1 (ja) |
PT (1) | PT1007654E (ja) |
WO (1) | WO1998037186A1 (ja) |
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US7416847B1 (en) | 2008-08-26 |
EP1007654B1 (en) | 2005-06-08 |
GB9703369D0 (en) | 1997-04-09 |
ATE297462T1 (de) | 2005-06-15 |
IL131435A (en) | 2008-06-05 |
NO322653B1 (no) | 2006-11-13 |
DK1007654T3 (da) | 2005-09-19 |
NO993953D0 (no) | 1999-08-17 |
NO993953L (no) | 1999-10-18 |
DE69830497D1 (de) | 2005-07-14 |
CA2282497C (en) | 2007-04-24 |
DE1007654T1 (de) | 2000-10-05 |
IL131435A0 (en) | 2001-01-28 |
PT1007654E (pt) | 2005-08-31 |
EP1007654A1 (en) | 2000-06-14 |
AU6108698A (en) | 1998-09-09 |
CA2282497A1 (en) | 1998-08-27 |
ES2241117T3 (es) | 2005-10-16 |
JP4644321B2 (ja) | 2011-03-02 |
DE69830497T2 (de) | 2006-03-16 |
WO1998037186A1 (en) | 1998-08-27 |
AU744791B2 (en) | 2002-03-07 |
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