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    Method of obtaining gene product through the generation of transgenic animals
    1.
    发明授权
    Method of obtaining gene product through the generation of transgenic animals 失效
    通过产生转基因动物获得基因产物的方法

    公开(公告)号：US4870009A

    公开(公告)日：1989-09-26

    申请号：US561644

    申请日：1983-12-15

    申请人： Ronald M. Evans ,  Richard D. Palmiter ,  Ralph L. Brinster

    发明人： Ronald M. Evans ,  Richard D. Palmiter ,  Ralph L. Brinster

    IPC分类号： C07K14/61 ,  C12N15/16 ,  C12N15/85 ,  C12N15/89

    CPC分类号： C07K14/61 ,  C12N15/85 ,  C12N15/89

    摘要： Mammalian genes that encode hormones are cloned and linked to strong promoter DNA sequences. The linked sequences are inserted in plasmids for amplification in prokaryotic cells, and multiple copies of the linked sequences are excised therefrom. Linked sequences are subsequently microinjected into fertilized eggs and the fertilized eggs are implanted into pseudo-pregnent females of the same species. As a result, transgenic animals are born having the linked sequences incorporated into their genomes and expressing the gene-encoded hormone. Because multiple copies of the linked sequences are frequently inserted and because production of the hormone is not limited to certain organs, as is the case with most endogenous hormones, the transgenic animals produce substantial amounts of the hormone. Hormone can be harvested from the living animal (and from its hormone-producing progeny) by extracting fluid, such as blood serum or ascites fluid, on a regular basis.

    摘要翻译： 编码激素的哺乳动物基因被克隆并与强启动子DNA序列连接。 将连接的序列插入质粒用于在原核细胞中扩增，并从其中切除连接序列的多个拷贝。 随后将连锁序列显微注射到受精卵中，将受精卵植入相同物种的假怀孕女性。 结果，转基因动物出生后将连锁序列并入其基因组并表达基因编码的激素。 由于连锁序列的多个拷贝经常插入，因为激素的产生不限于某些器官，如大多数内源激素的情况，转基因动物产生大量的激素。 通过定期地提取液体，如血清或腹水，可以从活的动物（以及来自其激素产生的后代）中收获激素。

    Synthesis of functional human hemoglobin and other proteins in erythroid tissues of transgenic animals
    2.
    发明授权
    Synthesis of functional human hemoglobin and other proteins in erythroid tissues of transgenic animals 失效
    在转基因动物的红细胞组织中合成功能性人血红蛋白和其他蛋白质

    公开(公告)号：US5766884A

    公开(公告)日：1998-06-16

    申请号：US466443

    申请日：1995-06-06

    申请人： Tim M. Townes ,  Thomas M. Ryan ,  Richard D. Palmiter ,  Ralph L. Brinster ,  Richard R. Behringer

    发明人： Tim M. Townes ,  Thomas M. Ryan ,  Richard D. Palmiter ,  Ralph L. Brinster ,  Richard R. Behringer

    IPC分类号： A01K67/027 ,  A61K38/00 ,  A61K48/00 ,  C07K14/805 ,  C12N15/85 ,  C12N15/00 ,  C12N15/79

    CPC分类号： C12N15/8509 ,  A01K67/0275 ,  A01K67/0278 ,  A61K48/00 ,  C07K14/805 ,  C12N15/85 ,  A01K2207/15 ,  A01K2217/00 ,  A01K2217/05 ,  A01K2227/105 ,  A01K2227/108 ,  A01K2267/01 ,  A01K2267/0306 ,  A01K2267/0381 ,  A61K38/00 ,  C12N2830/00 ,  C12N2830/008 ,  C12N2830/85 ,  Y10S435/948

    摘要： The present invention relates to the synthesis of functional human hemoglobin and other proteins in erythroid tissues of transgenic non-human animals and erythroid cell lines. It is based on the discovery that two of the five hypersensitivity sites of the .beta.-globin locus are sufficient to result in high level expression of human .alpha.- or .beta.-globin transgenes. The present invention also provides for novel recombinant nucleic acid vectors which may be used to produce .alpha.-globin as well as other proteins of interest in quantity in the red blood cells of transgenic animals or cell cultures of erythroid lineage. The vectors of the invention comprise at least one of the major DNase I hypersensitivity sites associated with the .beta.-globin locus together with a gene of interest. In a specific embodiment of the invention, a vector which comprises two DNase I hypersensitivity sites together with the human .alpha.-globin gene is used to create transgenic animals which produce human .alpha.-globin protein in erythroid tissues, including red blood cells. In a preferred specific embodiment of the invention, transgenic animals are created which comprise human .alpha.-globin and .beta.-globin genes, each under the transcriptional influence of two .beta.-globin locus DNase hypersensitivity sites; these transgenic animals express human hemoglobin in their erythroid tissues, and can be used to produce human hemoglobin in quantity.

    摘要翻译： 本发明涉及转基因非人动物和红系细胞系的红细胞组织中功能性人血红蛋白和其他蛋白质的合成。 基于以下发现：β-珠蛋白基因座的五个超敏性位点中的两个足以导致人α或β-珠蛋白转基因的高水平表达。 本发明还提供新的重组核酸载体，其可用于在转基因动物或红细胞谱系的细胞培养物的红细胞中产生量的α-珠蛋白以及其它目标蛋白。 本发明的载体包含与β-球蛋白基因座相关联的主要DNA酶I超敏反应位点与感兴趣的基因中的至少一种。 在本发明的一个具体实施方案中，使用包含两个DNA酶I超敏反应位点与人α-珠蛋白基因一起的载体来产生在包括红细胞在内的红细胞组织中产生人α-珠蛋白的转基因动物。 在本发明的优选具体实施方案中，产生包含人α-珠蛋白和β-珠蛋白基因的转基因动物，每个基因在两个β-球蛋白基因座DNA酶超敏位点的转录影响下; 这些转基因动物在其红细胞组织中表达人血红蛋白，并且可用于量产生人血红蛋白。

    Synthesis of functional human hemoglobin and other proteins in erythroid tissues of transgenic animals
    3.
    发明授权
    Synthesis of functional human hemoglobin and other proteins in erythroid tissues of transgenic animals 失效
    在转基因动物的红细胞组织中合成功能性人血红蛋白和其他蛋白质

    公开(公告)号：US5602306A

    公开(公告)日：1997-02-11

    申请号：US275313

    申请日：1994-07-13

    申请人： Tim M. Townes ,  Thomas M. Ryan ,  Richard D. Palmiter ,  Ralph L. Brinster ,  Richard R. Behringer

    发明人： Tim M. Townes ,  Thomas M. Ryan ,  Richard D. Palmiter ,  Ralph L. Brinster ,  Richard R. Behringer

    IPC分类号： A01K67/027 ,  A61K38/00 ,  A61K48/00 ,  C07K14/805 ,  C12N15/85 ,  C12N15/00

    CPC分类号： C12N15/8509 ,  A01K67/0275 ,  A01K67/0278 ,  A61K48/00 ,  C07K14/805 ,  C12N15/85 ,  A01K2207/15 ,  A01K2217/00 ,  A01K2217/05 ,  A01K2227/105 ,  A01K2227/108 ,  A01K2267/01 ,  A01K2267/0306 ,  A01K2267/0381 ,  A61K38/00 ,  C12N2830/00 ,  C12N2830/008 ,  C12N2830/85 ,  Y10S435/948

    摘要： The present invention relates to the synthesis of functional human hemoglobin and other proteins in erythroid tissues of transgenic non-human animals and erythroid cell lines. It is based on the discovery that two of the five hypersensitivity sites of the .beta.-globin locus are sufficient to result in high level expression of human .alpha.- or .beta.-globin transgenes. The present invention also provides for novel recombinant nucleic acid vectors which may be used to produce .alpha.-globin as well as other proteins of interest in quantity in the red blood cells of transgenic animals or cell cultures of erythroid lineage. The vectors of the invention comprise at least one of the major DNase I hypersensitivity sites associated with the .beta.-globin locus together with a gene of interest. In a specific embodiment of the invention, a vector which comprises two DNase I hypersensitivity sites together with the human .alpha.-globin gene is used to create transgenic animals which produce human .alpha.-globin protein in erythroid tissues, including red blood cells. In a preferred specific embodiment of the invention, transgenic animals are created which comprise human .alpha.-globin and .beta.-globin genes, each under the transcriptional influence of two .beta.-globin locus DNase hypersensitivity sites; these transgenic animals express human hemoglobin in their erythroid tissues, and can be used to produce human hemoglobin in quantity.

    摘要翻译： 本发明涉及转基因非人动物和红系细胞系的红细胞组织中功能性人血红蛋白和其他蛋白质的合成。 基于以下发现：β-珠蛋白基因座的五个超敏性位点中的两个足以导致人α或β-珠蛋白转基因的高水平表达。 本发明还提供新的重组核酸载体，其可用于在转基因动物或红细胞谱系的细胞培养物的红细胞中产生量的α-珠蛋白以及其它目标蛋白。 本发明的载体包含与β-球蛋白基因座相关联的主要DNA酶I超敏反应位点与感兴趣的基因中的至少一种。 在本发明的一个具体实施方案中，使用包含两个DNA酶I超敏反应位点与人α-珠蛋白基因一起的载体来产生在包括红细胞在内的红细胞组织中产生人α-珠蛋白的转基因动物。 在本发明的优选具体实施方案中，产生包含人α-珠蛋白和β-珠蛋白基因的转基因动物，每个基因在两个β-球蛋白基因座DNA酶超敏位点的转录影响下; 这些转基因动物在其红细胞组织中表达人血红蛋白，并且可用于量产生人血红蛋白。

    Control of DNA sequence transcription
    4.
    发明授权
    Control of DNA sequence transcription 失效
    控制DNA序列转录

    公开(公告)号：US4579821A

    公开(公告)日：1986-04-01

    申请号：US324111

    申请日：1981-11-23

    申请人： Richard D. Palmiter ,  Ralph L. Brinster

    发明人： Richard D. Palmiter ,  Ralph L. Brinster

    IPC分类号： C12N15/00 ,  C12N1/00 ,  C12N5/00 ,  C12P21/00

    CPC分类号： C12N15/00

    摘要： The transcription of DNA sequences in living cells is subjected to external regulation by incorporation of promoter/regulator DNA sequences responsive to metals and/or steroids. More particularly, regulation of the transcription of selected exogenous DNA sequences incorporated into eukaryotic host cells is facilitated by operative association (e.g., fusion) of the selected sequence to a promoter/regulator DNA sequence which is positively or negatively responsive to environmental variation in the concentration of heavy metal ions and/or steroid hormones. As an example, a structural gene for thymidine kinase from herpes simplex virus, fused to the promoter/regulator DNA sequence of a mouse metallothionein-I gene and incorporated on a suitable DNA plasmid vector, is introduced into mouse embryos and stably incorporated therein. Gene expression in differentiated cells of adult mice resulting from the embryos is subsequently regulatable by administration of heavy metals such as cadmium or steroid hormones such as the glucocorticoid, dexamethasone.

    摘要翻译： 通过掺入对金属和/或类固醇有响应的启动子/调控DNA序列，对活细胞中DNA序列的转录进行外部调控。 更具体地，通过将​​所选择的序列与启动子/调节子DNA序列的有效关联（例如，融合）促进掺入真核宿主细胞中的选择的外源DNA序列的转录调节，所述启动子/调节物DNA序列对于浓度的环境变化是正或负反应的 的重金属离子和/或类固醇激素。 作为例子，将来自单纯疱疹病毒的胸腺嘧啶激酶的结构基因融合到小鼠金属硫蛋白-I基因的启动子/调节子DNA序列上，并掺入合适的DNA质粒载体中，导入小鼠胚胎并稳定地并入其中。 通过施用重金属如镉或类固醇激素如糖皮质激素，地塞米松，随后可以调节由胚胎产生的成年小鼠的分化细胞中的基因表达。

    Nucleic acid vectors comprising DNase I hypersensitive sites
    6.
    发明授权
    Nucleic acid vectors comprising DNase I hypersensitive sites 失效
    包含DNase I过敏位点的核酸载体

    公开(公告)号：US6022736A

    公开(公告)日：2000-02-08

    申请号：US471483

    申请日：1995-06-06

    申请人： Tim M. Townes ,  Thomas M. Ryan ,  Richard D. Palmiter ,  Ralph L. Brinster ,  Richard R. Behringer

    发明人： Tim M. Townes ,  Thomas M. Ryan ,  Richard D. Palmiter ,  Ralph L. Brinster ,  Richard R. Behringer

    IPC分类号： A01K67/027 ,  A61K38/00 ,  A61K48/00 ,  C07K14/805 ,  C12N15/85 ,  C12N15/63 ,  C12N5/00 ,  C12N15/00 ,  C12N15/09

    CPC分类号： C12N15/8509 ,  A01K67/0275 ,  A01K67/0278 ,  A61K48/00 ,  C07K14/805 ,  C12N15/85 ,  A01K2207/15 ,  A01K2217/00 ,  A01K2217/05 ,  A01K2227/105 ,  A01K2227/108 ,  A01K2267/01 ,  A01K2267/0306 ,  A01K2267/0381 ,  A61K38/00 ,  C12N2830/00 ,  C12N2830/008 ,  C12N2830/85 ,  Y10S435/948

    摘要： The present invention provides novel recombinant nucleic acid vectors which may be used to produce .alpha.-globin as well as other proteins of interest in quantity in the red blood cells of transgenic animals or cell cultures of erythroid lineage. The present invention also provides for the transgenic animals which contain these recombinant nucleic acid vectors. The vectors of the invention comprise at least one of the major DNase I hypersensitivity sites associated with the .beta.-globin locus together with a gene of interest. According to various embodiments of the invention, the vectors may be used to create transgenic animals or to transfect cells in culture. In a specific embodiment of the invention, a vector which comprises two DNase I hypersensitivity sites together with the human .alpha.-globin gene is used to create transgenic animals which produce human .alpha.-globin protein in erythroid tissues, including red blood cells. In a preferred specific embodiment of the invention, transgenic animals are created which comprise human .alpha.-globin and .beta.-globin genes, each under the transcriptional influence of two .beta.-globin locus DNase hypersensitivity sites; these transgenic animals express human hemoglobin in their erythroid tissues, and can be used to produce human hemoglobin in quantity.

    摘要翻译： 本发明提供新的重组核酸载体，其可用于在转基因动物或红细胞谱系的细胞培养物的红细胞中产生量的α-珠蛋白以及其它目标蛋白。 本发明还提供含有这些重组核酸载体的转基因动物。 本发明的载体包含与β-球蛋白基因座相关联的主要DNA酶I超敏反应位点与感兴趣的基因中的至少一种。 根据本发明的各种实施方案，载体可用于产生转基因动物或转染培养物中的细胞。 在本发明的一个具体实施方案中，使用包含两个DNA酶I超敏反应位点与人α-珠蛋白基因一起的载体来产生在包括红细胞在内的红细胞组织中产生人α-珠蛋白的转基因动物。 在本发明的优选具体实施方案中，产生包含人α-珠蛋白和β-珠蛋白基因的转基因动物，每个基因在两个β-球蛋白基因座DNA酶超敏位点的转录影响下; 这些转基因动物在其红细胞组织中表达人血红蛋白，并且可用于量产生人血红蛋白。