JPH11511009A - 細胞への核酸運搬のためのプラスミドおよび使用方法 - Google Patents
細胞への核酸運搬のためのプラスミドおよび使用方法Info
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- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
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- 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
- C12N15/70—Vectors or expression systems specially adapted for E. coli
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
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- C12N2800/00—Nucleic acids vectors
- C12N2800/30—Vector systems comprising sequences for excision in presence of a recombinase, e.g. loxP or FRT
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- C12N2820/00—Vectors comprising a special origin of replication system
- C12N2820/002—Vectors comprising a special origin of replication system inducible or controllable
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- C12N2820/00—Vectors comprising a special origin of replication system
- C12N2820/55—Vectors comprising a special origin of replication system from bacteria
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- C12N2830/00—Vector systems having a special element relevant for transcription
- C12N2830/001—Vector systems having a special element relevant for transcription controllable enhancer/promoter combination
- C12N2830/002—Vector systems having a special element relevant for transcription controllable enhancer/promoter combination inducible enhancer/promoter combination, e.g. hypoxia, iron, transcription factor
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- C12N2830/00—Vector systems having a special element relevant for transcription
- C12N2830/001—Vector systems having a special element relevant for transcription controllable enhancer/promoter combination
- C12N2830/005—Vector systems having a special element relevant for transcription controllable enhancer/promoter combination repressible enhancer/promoter combination, e.g. KRAB
- C12N2830/006—Vector systems having a special element relevant for transcription controllable enhancer/promoter combination repressible enhancer/promoter combination, e.g. KRAB tet repressible
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- C12N2830/00—Vector systems having a special element relevant for transcription
- C12N2830/36—Vector systems having a special element relevant for transcription being a transcription termination element
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- C12N2830/00—Vector systems having a special element relevant for transcription
- C12N2830/55—Vector systems having a special element relevant for transcription from bacteria
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N2840/00—Vectors comprising a special translation-regulating system
- C12N2840/20—Vectors comprising a special translation-regulating system translation of more than one cistron
Abstract
Description
Claims (1)
- 【特許請求の範囲】 1.ヒト細胞へ核酸配列を運搬するプラスミドであって、 複製開始配列を含みプラスミドコピー数を制御する5’隣接領域、 プラスミド保有細胞の選択を制御し前記5’隣接領域に対してさまざまに位置 している3’隣接領域であって、抗生物質耐性をコードする核酸配列を含む前記 3’隣接領域、 前記3’隣接領域に対して前記5’隣接領域を接続するリンカー領域であって 、核酸カセット挿入のためのマルチクローニング部位要素、および前記抗生物質 耐性配列の転写が前記核酸カセットにまで転写されないように確実にする転写終 止要素を含む前記リンカー領域、 を含む前記プラスミド。 2.前記5’隣接領域の複製開始点に隣接する複数の核酸制限酵素部位、前記 3’隣接領域の前記抗生物質耐性核酸配列、前記リンカー領域および前記転写終 止配列をさらに含む請求項1記載のプラスミド。 3.前記プラスミドが前記核酸制限酵素部位中に挿入された機能要素をさらに 含み、前記機能要素が分配座またはマルチマー分割座からなる群から選択される 請求項2記載のプラスミド。 4.前記複製開始点がプラスミドpMM1由来であり、前記抗生物質耐性核酸 配列がneo遺伝子であり、前記マルチクローニング部位がpBluescri pt KS+由来のポリリンカー配列であり、および前記転写終止配列がプラス ミドpKK223−3由来のRRNBT1T2である請求項1記載のプラスミド。 5.細胞へ核酸カセットを運搬するためにプラスミドを使用する方法であって 、請求項1、2、3または4のプラスミドに前記細胞を十分時間接触させ、前記 細胞を形質転換させる工程を含み、請求項1、2、3または4のプラスミドが前 記核酸カセットをさらに含む前記方法。 6.細胞へ核酸配列を運搬するプラスミドであって、 複製開始配列を含みプラスミドコピー数を制御する5’隣接領域、 プラスミド保有細胞の選択を制御し前記5’隣接領域に対してさまざまに位置 している3’隣接領域であって、抗生物質耐性をコードする核酸配列を含みかつ 複数の核酸制限酵素配列に隣接している前記3’隣接領域、 前記5’隣接領域と前記3’隣接領域の間にあるリンカー領域であって、核酸 カセット挿入のためのマルチクローニング部位要素、前記抗生物質耐性配列の転 写が前記核酸カセットにまで確実に転写されないようにする転写終止要素、およ び染色体オペロンを脱抑制できるオペレータ配列からなる前記リンカー領域、 を含む前記プラスミド。 7.前記オペレータ配列が前記5’隣接領域に対して5’にかつ前記3’隣接 領域に対して5’に位置し、それが多様に配向されている請求項6記載のプラス ミド。 8.前記オペレータ配列が前記5’隣接領域に対して3’にかつ前記リンカー 領域に対して5’に位置する請求項6記載のプラスミド。 9.前記プラスミドが複数の前記オペレータ配列をさらに含む請求項6記載の プラスミド。 10.前記オペレータ配列が前記5’隣接領域に対して5’にかつ前記3’隣接 領域に対して3’に位置しており、かつ前記5’隣接領域に対して3’にかつ前 記リンカー領域に対して5’に位置している請求項9記載のプラスミド。 11.細胞へ核酸カセットを運搬するためにプラスミドを使用する方法であって 、請求項6、7、8、9または10に記載のプラスミドに前記細胞を十分時間接 触させ、前記細胞を形質転換させる工程を含み、請求項6、7、8、9または1 0に記載のプラスミドが前記核酸カセットをさらに含む前記方法。 12.細胞へ核酸配列を運搬するプラスミドであって、 複製開始配列を含みプラスミドコピー数を制御する5’隣接領域、 前記5’隣接領域に対して3’のリンカー領域であって、核酸カセット挿入の ためのマルチクローニング部位要素を含む前記リンカー領域、および 染色体オペロンの脱抑制が可能なオペレータ配列、 を含む前記プラスミド。 13.前記オペレータ配列が前記3’リンカー領域に対して3’に位置しかつ前 記5’隣接領域に対して5’に位置する請求項12記載のプラスミド。 14.前記オペレータ配列が前記5’隣接領域に対して3’にかつ前記リンカー 領域に対して5’に位置する請求項12記載のプラスミド。 15.前記プラスミドが前記リンカー領域に対して3’の3’隣接領域をさらに 含み、前記3’隣接領域が転写終止要素および少なくとも1個の核酸制限酵素部 位を含む請求項12記載のプラスミド。 16.複数のオペレータ配列をさらに含む請求項12記載のプラスミド。 17.前記オペレータ配列が前記5’隣接領域に対して5’にかつ前記3’隣接 領域に対して3’に位置しており、かつ前記5’隣接領域に対して3’にかつ前 記リンカー領域に対して5’に位置している請求項16記載のプラスミド。 18.細胞への核酸カセット運搬のためにプラスミドを使用する方法であって、 請求項12、13、14、15、16または17に記載のプラスミドに前記細胞 を十分時間接触させ、前記細胞を形質転換させる工程を含み、請求項12、13 、14、15、16または17に記載のプラスミドは前記核酸カセットをさらに 含む前記方法。 19.プラスミド選択方法であって、以下の工程: 第1の細胞を複数のプラスミドに十分時間接触させ前記細胞を形質転換させる ことによって、プラスミド群からプラスミドを選択し、ここで前記プラスミドは 、請求項6、7、8、9または10に記載のプラスミドでありさらに前記核酸カ セットを含み、 前記第1の細胞を適切に形質転換させるプラスミドを選択し、 隣接核酸制限酵素配列のところで前記選択されたプラスミドから抗生物質耐性 配列を除去し、 前記選択されたプラスミドを再環状化し、 第2の細胞を前記選択されたプラスミドに十分時間接触させ前記第2の細胞を 形質転換させ、ここで前記第2の細胞は、前記細胞の染色体中に染色体オペロン 制御下に取り込まれた少なくとも1個の抗生物質耐性遺伝子を保有し、前記オペ ロンが前記選択されたプラスミド上のオペレータ配列によって脱抑制可能であり 、 形質転換された抗生物質耐性の第2の細胞を選択し、そして 前記選択された第2の細胞から前記選択されたプラスミドを単離する、 を含む前記方法。 20.プラスミド選択方法であって、以下の工程: 細胞をプラスミドに十分時間接触させ前記細胞を形質転換させ、ここで前記プ ラスミドは、請求項12、13、14、15、16または17に記載のプラスミ ドであり前記核酸カセットをさらに含み、前記細胞は、前記細胞の染色体中に染 色体オペロンの制御下に取り込まれた少なくとも1個の抗生物質耐性遺伝子を保 有し、前記オペロンは、前記プラスミド上の前記オペレータ配列による脱抑制が 可能であり、 抗生物質耐性の前記形質転換された細胞を選択し、 前記選択された細胞から前記プラスミドを単離する、 を含む前記方法。 21.プラスミド選択のための系であって、 前記核酸カセットを含む請求項6、7、8、9または10に記載のプラスミド 、 前記核酸カセットを含む請求項12、13、14、15、16または17に記 載のプラスミド、および 前記細胞の染色体中に染色体オペロンの制御下に取り込まれた少なくとも1個 の抗生物質耐性遺伝子を保有する細胞であって、前記オペロンは、前記プラスミ ド上の前記オペレータ配列による脱抑制が可能である前記細胞、 を含む前記系。 22.pVC0396と命名されたプラスミド。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US08/484,723 US5763270A (en) | 1995-06-07 | 1995-06-07 | Plasmid for delivery of nucleic acids to cells and methods of use |
US08/484,723 | 1995-06-07 | ||
PCT/US1996/005680 WO1996040952A1 (en) | 1995-06-07 | 1996-04-23 | Plasmid for delivery of nucleic acids to cells and methods of use |
Publications (2)
Publication Number | Publication Date |
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JPH11511009A true JPH11511009A (ja) | 1999-09-28 |
JP4335309B2 JP4335309B2 (ja) | 2009-09-30 |
Family
ID=23925336
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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JP50048597A Expired - Fee Related JP4335309B2 (ja) | 1995-06-07 | 1996-04-23 | 細胞への核酸運搬のためのプラスミドおよび使用方法 |
Country Status (9)
Country | Link |
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US (2) | US5763270A (ja) |
EP (1) | EP0832264B1 (ja) |
JP (1) | JP4335309B2 (ja) |
AT (1) | ATE323169T1 (ja) |
AU (1) | AU711187B2 (ja) |
CA (1) | CA2222601C (ja) |
DE (1) | DE69636032T2 (ja) |
PT (1) | PT832264E (ja) |
WO (1) | WO1996040952A1 (ja) |
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US5019506A (en) * | 1986-05-21 | 1991-05-28 | University College, Cork | Plasmid and uses thereof |
HU197595B (en) * | 1987-09-16 | 1989-04-28 | Richter Gedeon Vegyeszet | Process for producing expression vectors |
US4956288A (en) * | 1988-04-22 | 1990-09-11 | Biogen, Inc. | Method for producing cells containing stably integrated foreign DNA at a high copy number, the cells produced by this method, and the use of these cells to produce the polypeptides coded for by the foreign DNA |
DE3819463A1 (de) * | 1988-06-08 | 1989-12-14 | Behringwerke Ag | Expressionsvektoren zur herstellung unfusionierter proteine in mikroorganismen |
US5492890A (en) * | 1990-12-03 | 1996-02-20 | The Scripps Research Institute | Polypeptides for promoting cell attachment |
US5308760A (en) * | 1992-01-10 | 1994-05-03 | Washington Research Foundation | Crystal proteins of Bacillus thuringiensis, genes encoding them, and host expressing them |
GB9312070D0 (en) * | 1993-06-11 | 1993-07-28 | Thrombosis Res Inst | Untranslated exon-1 sequences of eukaryotic genes as promoters |
US5763270A (en) * | 1995-06-07 | 1998-06-09 | Genemedicine, Inc. | Plasmid for delivery of nucleic acids to cells and methods of use |
GB9518395D0 (en) * | 1995-09-08 | 1995-11-08 | Therexsys Ltd | Plasmid stabilization |
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AU711187B2 (en) | 1999-10-07 |
CA2222601A1 (en) | 1996-12-19 |
US5763270A (en) | 1998-06-09 |
JP4335309B2 (ja) | 2009-09-30 |
AU5788096A (en) | 1996-12-30 |
WO1996040952A1 (en) | 1996-12-19 |
US6103470A (en) | 2000-08-15 |
ATE323169T1 (de) | 2006-04-15 |
DE69636032D1 (de) | 2006-05-24 |
DE69636032T2 (de) | 2006-11-23 |
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