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    Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials
    1.
    发明授权
    Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials 失效
    将DNA插入真核细胞并生产蛋白质材料的方法

    公开(公告)号：US4399216A

    公开(公告)日：1983-08-16

    申请号：US124513

    申请日：1980-02-25

    申请人： Richard Axel ,  Michael H. Wigler ,  Saul J. Silverstein

    发明人： Richard Axel ,  Michael H. Wigler ,  Saul J. Silverstein

    IPC分类号： C12N5/00 ,  C12N15/00 ,  C12N15/85 ,  C12N15/87 ,  C12P21/00 ,  C12P21/02 ,  C12Q1/02 ,  C12Q1/04 ,  C12Q1/68

    CPC分类号： C12N15/87 ,  C12N15/85 ,  C12P21/02 ,  Y10S435/811 ,  Y10S435/948

    摘要： The present invention relates to processes for inserting DNA into eucaryotic cells, particularly DNA which includes a gene or genes coding for desired proteinaceous materials for which no selective criteria exist. The insertion of such DNA molecules is accomplished by cotransforming eucaryotic cells with such DNA together with a second DNA which corresponds to a gene coding for a selectable marker.The invention further relates to processes for inserting into eucaryotic cells a multiplicity of DNA molecules including genes coding for desired proteinaceous materials by cotransformation with the desired genes and with amplifiable genes for a dominant selectable marker in the presence of successively higher amounts of an inhibitor. Alternatively, the insertion of multiple copies of desired genes is accomplished by transformation using DNA molecules formed by ligating a DNA molecule including the desired gene to a DNA molecule which includes an amplifiable gene coding for a dominant selectable phenotype such as a gene associated with resistance to a drug in the presence of successively higher amounts of an agent such as a drug against which the gene confers resistance so that only those eucaryotic cells into which multiple copies of the amplifiable gene have been inserted survive.

    摘要翻译： 本发明涉及将DNA插入真核细胞的方法，特别是包括编码没有选择性标准的所需蛋白质材料的基因或基因的DNA。 这样的DNA分子的插入通过与这样的DNA共转化真核细胞以及对应于编码选择标记的基因的第二DNA来实现。 本发明还涉及在真核细胞中插入多个DNA分子的方法，所述多个DNA分子包括编码所需蛋白质材料的基因，通过与期望的基因共转化，并且在连续更高量的抑制剂存在下，具有用于显性选择标记的可扩增基因。 或者，通过使用通过将包含所需基因的DNA分子连接到DNA分子而形成的DNA分子进行转化来实现，所述DNA分子包括编码显性可选择表型的可扩增基因，例如与抗性相关的基因 在连续更多量的药物存在下的药物，所述药物如基因赋予抗性的药物，使得只有插入了可扩增基因的多个拷贝的真核细胞才能存活。

    DNA construct for producing proteinaceous materials in eucaryotic cells
    2.
    发明授权
    DNA construct for producing proteinaceous materials in eucaryotic cells 失效
    用于在真核细胞中产生蛋白质物质的DNA构建体

    公开(公告)号：US06455275B1

    公开(公告)日：2002-09-24

    申请号：US08484136

    申请日：1995-06-07

    申请人： Richard Axel ,  Michael H. Wigler ,  Saul J. Silverstein

    发明人： Richard Axel ,  Michael H. Wigler ,  Saul J. Silverstein

    IPC分类号： C12P2102

    CPC分类号： C12N15/87 ,  C12N15/85 ,  C12P21/02

    摘要： The invention relates to processes for inserting into eucaryotic cells a multiplicity of DNA molecules which includes genes coding for desired proteinaceous materials. The insertion of multiple copies of desired genes is accomplished by cotransformation with the desired genes and with amplifiable genes for a dominant selectable marker in the presence of successively higher amounts of an inhibitor. Alternatively, the insertion of multiple copies of desired genes is accomplished by transformation using DNA molecules formed by ligating a DNA molecule including the desired gene to a DNA molecule which includes an amplifiable gene coding for a dominant selectable phenotype such as a gene associated with resistance to a drug in the presence of successively higher amounts of an agent such as a drug against which the gene confers resistance so that only those eucaryotic cells into which multiple copies of the amplifiable gene have been inserted survive. Eucaryotic cells into which multiple copies of the amplifiable gene have been inserted additionally include multiple copies of the desired gene and may be used to produce multiple copies of proteinaceous molecules. In this way otherwise rare proteinaceous materials may be obtained in higher concentrations than are obtainable using conventional techniques.

    摘要翻译： 本发明涉及在真核细胞中插入多个DNA分子的方法，其包括编码所需蛋白质材料的基因。 所需基因的多个拷贝的插入通过与所需基因的共转化和用连续更高量的抑制剂存在的显性选择标记的可扩增基因来实现。 或者，通过使用通过将包含所需基因的DNA分子连接到DNA分子而形成的DNA分子进行转化来实现，所述DNA分子包括编码显性可选择表型的可扩增基因，例如与抗性相关的基因 在连续更多量的药物存在下的药物，所述药物如基因赋予抗性的药物，使得只有插入了可扩增基因的多个拷贝的真核细胞才能存活。 已插入多个拷贝的可扩增基因的真核细胞另外包含所需基因的多个拷贝，并可用于产生蛋白质分子的多个拷贝。 以这种方式，否则可以以比使用常规技术可获得的浓度更高的浓度获得稀有的蛋白质材料。

    Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials
    3.
    发明授权
    Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials 失效

    公开(公告)号：US5179017A

    公开(公告)日：1993-01-12

    申请号：US716915

    申请日：1991-06-18

    申请人： Richard Axel ,  Michael H. Wigler ,  Saul J. Silverstein

    发明人： Richard Axel ,  Michael H. Wigler ,  Saul J. Silverstein

    IPC分类号： C12N15/85 ,  C12N15/87 ,  C12P21/02

    CPC分类号： C12N15/85 ,  C12N15/87 ,  C12P21/02

    摘要： The present invention relates to processes for inserting DNA into eucaryotic cells, particularly DNA which includes a gene or genes coding for desired proteinaceous materials for which no selective criteria exist. The insertion of such DNA molecules is accomplished by cotransforming eucaryotic cells with such DNA together with a second DNA which corresponds to a gene coding for a selectable marker.This invention also concerns processes for producing proteinaceous materials such as insulin, interferon protein, growth hormone and the like which involve cotransforming eucaryotic cells with DNA which codes for these proteinaceous materials, growing the cotransformed cells for production of the proteinaceous material and recovering the proteinaceous material so produced.The invention further relates to processes for inserting into eucaryotic cells a multiplicity of DNA molecules which includes genes coding for desired proteinaceous materials. The insertion of multiple copies of desired genes is accomplished by cotransformation with the desired genes and with amplifiable genes for a dominant selectable marker in the presence of successively higher amounts of an inhibitor. Alternatively, the insertion of multiple copies of desired genes is accomplished by transformation using DNA molecules formed by ligating a DNA molecule including the desired gene to a DNA molecule which includes an amplifiable gene coding for a dominant selectable phenotype such as a gene associated with resistance to a drug in the presence of successively higher amounts of an agent such as a drug against which the gene confers resistance so that only those eucaryotic cells into which multiple copies of the amplifiable gene have been inserted survive. Eucaryotic cells into which multiple copies of the amplifiable gene have been inserted additionally include multiple copies of the desired gene and may be used to produce multiple copies of proteinaceous molecules. In this way otherwise rare proteinaceous materials may be obtained in higher concentrations than are obtainable using conventional techniques.

    Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials
    4.
    发明授权
    Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials 失效

    公开(公告)号：US4634665A

    公开(公告)日：1987-01-06

    申请号：US522408

    申请日：1983-08-11

    申请人： Richard Axel ,  Michael H. Wigler ,  Saul J. Silverstein

    发明人： Richard Axel ,  Michael H. Wigler ,  Saul J. Silverstein

    IPC分类号： C12N15/85 ,  C12P21/02 ,  C12P21/00 ,  C12N5/00 ,  C12N15/00 ,  C12R1/91

    CPC分类号： C12N15/85 ,  C12P21/02 ,  Y10S435/811 ,  Y10S435/948

    摘要： The present invention relates to processes for inserting DNA into eucaryotic cells, particularly DNA which includes a gene or genes coding for desired proteinaceous materials for which no selective criteria exist. The insertion of such DNA molecules is accomplished by cotransforming eucaryotic cells with such DNA together with a second DNA which corresponds to a gene coding for a selectable marker.This invention also concerns processes for producing proteinaceous materials such as insulin, interferon protein, growth hormone and the like which involve cotransforming eucaryotic cells with DNA which codes for these proteinaceous materials, growing the contransformed cells for production of the proteinaceous material and recovering the proteinaceous material so produced.The invention further relates to processes for inserting into eucaryotic cells a multiplicity of DNA molecules which includes genes coding for desired proteinaceous materials. The insertion of multiple copies of desired genes is accomplished by cotransformation with the desired genes and with amplifiable genes for a dominant selectable marker in the presence of successively higher amounts of an inhibitor. Alternatively, the insertion of multiple copies of desired genes is accomplished by transformation using DNA molecules formed by ligating a DNA molecule including the desired gene to a DNA molecule which includes an amplifiable gene coding for a dominant selectable phenotype such as a gene associated with resistance to a drug in the presence of successively higher amounts of an agent such as a drug against which the gene confers resistance so that only those eucaryotic cells into which multiple copies of the amplifiable gene have been inserted survive. Eucaryotic cells into which multiple copies of the amplifiable gene have been inserted additionally include multiple copies of the desired gene and may be used to produce multiple copies of proteinaceous molecules. In this way otherwise rare proteinaceous materials may be obtained in higher concentrations than are obtainable using conventional techniques.

    VZV ORF29p protein-related compositions and methods
    8.
    发明授权
    VZV ORF29p protein-related compositions and methods 失效
    VZV ORF29p蛋白相关组合物和方法

    公开(公告)号：US06809182B2

    公开(公告)日：2004-10-26

    申请号：US09769699

    申请日：2001-01-25

    申请人： Saul J. Silverstein ,  Paula W. Annuziato ,  Anne A. Gershon ,  Octavian Lungu

    发明人： Saul J. Silverstein ,  Paula W. Annuziato ,  Anne A. Gershon ,  Octavian Lungu

    IPC分类号： C07K1400

    CPC分类号： C07K14/005 ,  A61K9/127 ,  A61K38/00 ,  A61K47/62 ,  A61K48/00 ,  C07K2319/00 ,  C12N2710/16722

    摘要： The present invention provides compositions of matter comprising 29p protein having bound thereto an agent whose delivery into a eukaryotic cell is desired. The present invention also provides a monoclonal antibody which specifically binds to 29p protein. The present invention further provides methods for delivering an agent into a eukaryotic cell, and methods for causing a eukaryotic cell to secrete a desired protein in the form of a fusion protein. The present invention further provides 29p protein-containing pharmaceutical compositions. The present invention still further provides nucleic acid molecules which hybridize to at least a portion of a nucleic acid molecule encoding 29p protein. Finally, the present invention provides methods for detecting the presence of, and quantitatively determining the amount of, a 29p protein-encoding nucleic acid molecule in a sample.

    摘要翻译： 本发明提供了包含29p蛋白质的物质组合物，其结合有期望其递送至真核细胞的试剂。 本发明还提供了与29p蛋白特异性结合的单克隆抗体。 本发明还提供了将试剂递送到真核细胞中的方法，以及用于使真核细胞分泌融合蛋白形式的所需蛋白质的方法。 本发明还提供了含29p蛋白质的药物组合物。 本发明还提供了与至少一部分编码29p蛋白的核酸分子杂交的核酸分子。 最后，本发明提供了检测样品中29p蛋白编码核酸分子的存在和定量确定量的方法。

    Detection of high oncogenic-risk papilloma virus in high grade cervical lesions and cancers by a PCR/ELISA assay
    9.
    发明授权
    Detection of high oncogenic-risk papilloma virus in high grade cervical lesions and cancers by a PCR/ELISA assay 失效
    通过PCR / ELISA测定检测高级宫颈病变和癌症中的高致癌风险乳头瘤病毒

    公开(公告)号：US5888724A

    公开(公告)日：1999-03-30

    申请号：US479777

    申请日：1995-06-07

    申请人： Saul J. Silverstein ,  Octavian Lungu ,  Thomas C. Wright ,  Ralph M. Richart

    发明人： Saul J. Silverstein ,  Octavian Lungu ,  Thomas C. Wright ,  Ralph M. Richart

    IPC分类号： C12Q1/68 ,  C12Q1/70 ,  G01N33/574 ,  C12P19/34

    CPC分类号： C12Q1/686 ,  C12Q1/6804 ,  C12Q1/708 ,  G01N33/57411

    摘要： This invention provides a method of detecting a high oncogenic-risk type human papillomavirus in a subject which comprises: obtaining from a subject a specimen containing cervical cells and treating the specimen so as to recover nucleic acid molecules present in the cervical cells; contacting the resulting nucleic acid molecules with multiple pairs of single-stranded labeled oligonucleotide primers capable of specifically hybridizing with a different high oncogenic-risk type of human papillomavirus; amplifying any nucleic acid molecules to which a pair of primers hybridizes so as to obtain a double-stranded amplification product and treating any double-stranded amplification product so as to obtain single-stranded nucleic acid molecules; contacting any resulting single-stranded nucleic acid molecules with multiple single-stranded labeled oligonucleotide probes which are capable of specifically hybridizing with such high oncogenic-risk types of human papillomavirus; contacting any resulting hybrids with a marked antibody capable of specifically forming a complex with the labeled probe, when the probe is present in such a complex; and detecting the presence of any resulting complexes, the presence thereof being indicative of the presence of a high oncogenic-risk type human papillomavirus in the initial specimen.

    摘要翻译： 本发明提供一种检测受试者中高致癌危险型人乳头瘤病毒的方法，包括：从受试者获得含有宫颈细胞的标本并处理该标本以回收存在于宫颈细胞中的核酸分子; 使得到的核酸分子与能够与不同的高致癌危险类型的人乳头瘤病毒特异性杂交的多对单链标记寡核苷酸引物接触; 扩增一对引物杂交的任何核酸分子，以获得双链扩增产物并处理任何双链扩增产物，从而获得单链核酸分子; 使任何得到的单链核酸分子与能够与这种高致癌风险类型的人乳头瘤病毒特异性杂交的多个单链标记的寡核苷酸探针接触; 当所述探针存在于所述复合物中时，使所得到的杂交体与能够与标记的探针特异形成复合物的标记抗体接触; 并检测任何所得复合物的存在，其存在表明初始样品中存在高致癌危险型人乳头瘤病毒。