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12 条记录 (耗时 0.072 秒)

    Co-expression of alpha and beta globins
    4.
    发明授权
    Co-expression of alpha and beta globins 失效

    公开(公告)号：US5798227A

    公开(公告)日：1998-08-25

    申请号：US446105

    申请日：1995-05-19

    申请人： Stephen J. Hoffman ,  Douglas L. Looker ,  Gary L. Stetler ,  Michael Wagenbach

    发明人： Stephen J. Hoffman ,  Douglas L. Looker ,  Gary L. Stetler ,  Michael Wagenbach

    IPC分类号： A61K38/16 ,  A61K38/00 ,  A61K47/48 ,  A61P7/08 ,  C07K20060101 ,  C07K1/113 ,  C07K14/805 ,  C12N15/00 ,  C12N15/09 ,  C12N15/12 ,  C12N15/81 ,  C12P21/02 ,  C12R1/19 ,  C12R1/91 ,  C12P21/06 ,  C12N1/20 ,  C12P21/04

    CPC分类号： C07K14/805 ,  A61K47/48307 ,  C12N15/81 ,  A61K38/00 ,  C07K2319/00 ,  C07K2319/50 ,  C07K2319/92

    摘要： The alpha subunits of hemoglobin, which in nature are formed as separate polypeptide chains which bind noncovalently to the beta subunits, are here provided in the form of the novel molecule di-alpha globin, a single polypeptide chain defined by connecting the two alpha subunits either directly via peptide bond or indirectly by a flexible amino acid or peptide linker. Di-alpha globin may be combined in vivo or in vitro with beta globin and heme to form hemoglobin. Di-alpha globin is expressed by recombinant DNA techniques. Di-beta globin may be similarly obtained. We further describe the production of tetrameric human hemoglobin and di-alpha/beta.sub.2 hemoglobin in the yeast Saccharomyces cerevisiae. The synthesis of the protein is directed by a synthetic promotor consisting of two functional parts, an upstream activator sequence (UAS) that confers inducible transcription by galactose from a consensus yeast transcriptional initiation site. The expression construct is designed such that translation is expected to initiate at the same position as the human wild-type genes for .alpha.- and .beta.-globin. Three different types of expression vectors have been used: (1) .alpha.-globin and .beta.-globin contained on two separate plasmids (pGS4688 and pGS4988) in a diploid yeast strain; (2) .alpha.-globin and .beta.-globin each contained on a single plasmid (pGS289 and pGS389) and expressed in either haploid or diploid strains; and (3) di-alpha-globin and beta globin contained on a single plasmid (pGS 3089)and expressed in haploid strains. Finally, we describe the co-expression of alpha and beta globin chains. The chains are folded together and combined intracellularly with heme to form active tetrameric hemoglobin. The hemoglobin may be recovered from the cells' soluble fraction. The invention thus obviates the need to express alpha and beta globin separately, solubilize, renature and purify them, and combine them in vitro with heme to obtain an artificial hemoglobin. By way of comparison, the separately expressed beta globin known in the art is deposited in inclusion bodies. Polycistronic co-expression of alpha and beta globins is particularly preferred.

    Method of determining a functional linker for fusing globin subunts
    6.
    发明授权
    Method of determining a functional linker for fusing globin subunts 失效
    确定用于融合珠蛋白的功能性连接体的方法

    公开(公告)号：US06274331B1

    公开(公告)日：2001-08-14

    申请号：US08444915

    申请日：1995-05-19

    申请人： Douglas L. Looker ,  Gary L. Stetler

    发明人： Douglas L. Looker ,  Gary L. Stetler

    IPC分类号： G01N3353

    CPC分类号： C07K14/805 ,  A61K38/00 ,  A61K47/6445 ,  C07K2319/00 ,  C07K2319/50 ,  C07K2319/92 ,  C12N15/81

    摘要： A functional linker for a polypeptide in which two alpha or beta globin-like domains are genetically fused is determined by screening a library of genetically fused polypeptides, in which the linker region is varied, for the ability to participate in the formation of hemoglobin-like protein, as measured by the protein's response to carbon monoxide. In a preferred embodiment, cells expressing the protein turn red as a result of carbon monoxide pressure.

    摘要翻译： 通过筛选遗传融合多肽的文库，测定其中两个α或β球蛋白样结构域经遗传融合的功能性连接体，其中连接区不同，参与血红蛋白样形成的能力 蛋白质，通过蛋白质对一氧化碳的反应测量。 在优选的实施方案中，由于一氧化碳压力，表达蛋白质的细胞变红。

    Peptide inhibitors of tax-dependent transcription
    8.
    发明授权
    Peptide inhibitors of tax-dependent transcription 失效
    纳税依赖性转录肽抑制剂

    公开(公告)号：US5717058A

    公开(公告)日：1998-02-10

    申请号：US199508

    申请日：1994-02-18

    申请人： Maura-Ann H. Matthews ,  Gary L. Stetler ,  Spencer J. Anthony-Cahill ,  David C. Anderson

    发明人： Maura-Ann H. Matthews ,  Gary L. Stetler ,  Spencer J. Anthony-Cahill ,  David C. Anderson

    IPC分类号： A61K38/00 ,  A61K48/00 ,  C07K14/02 ,  C07K14/15 ,  C12N15/67 ,  C07K14/155 ,  C07K4/02 ,  C07K5/00 ,  C07K7/00

    CPC分类号： C07K14/005 ,  C12N15/67 ,  A01K2217/05 ,  A61K38/00 ,  A61K48/00 ,  C07K2319/00 ,  C12N2730/10122 ,  C12N2740/14022

    摘要： The present invention relates to regulators of cellular gene transcription, particularly inhibitors of cellular gene transactivating factors and in particular to inhibition of gene transcription in a viral host cell that is subject to regulation by proteins or factors that originate from a virus as well as conjugates or fusion products of the inhibitors and internalization molecules, pharmaceutical compositions that can be used to alleviate or prevent the manifestation of disease states that are the result of unregulated DNA transcription as a result of transactivation, methods of treating diseases that are caused or exacerbated by the presence of transactivating factors, and regulated gene therapy to achieve long term drug delivery of the inhibitors of the present invention. This invention can be applied both to cells with genetic abnormalities or to cells infected with a virus. Preferably, at least one protein of the protein-protein interactions is a transactivating factor.

    摘要翻译： 本发明涉及细胞基因转录的调节剂，特别是细胞基因反式激活因子的抑制剂，特别是抑制病毒宿主细胞中的基因转录，所述基因转录受来自病毒的蛋白质或因子的调节，以及缀合物或 抑制剂和内化分子的融合产物，可用于缓解或预防作为反式激活导致的不受调节的DNA转录的结果的疾病状态的表现的药物组合物，治疗由于存在引起或加剧的疾病的方法 的转录因子和调节基因治疗以实现本发明的抑制剂的长期药物递送。 本发明可以应用于具有遗传异常的细胞或用于感染病毒的细胞。 优选地，蛋白质 - 蛋白质相互作用的至少一种蛋白质是反式激活因子。

    Recombinant methods for production of serine protease inhibitors and DNA sequences useful for same
    10.
    发明授权
    Recombinant methods for production of serine protease inhibitors and DNA sequences useful for same 失效
    用于生产丝氨酸蛋白酶抑制剂和可用于其的DNA序列的重组方法

    公开(公告)号：US6132990A

    公开(公告)日：2000-10-17

    申请号：US712354

    申请日：1991-06-07

    申请人： Pradip K. Bandyopadhyay ,  Stephen P. Eisenberg ,  Gary L. Stetler ,  Robert C. Thompson

    发明人： Pradip K. Bandyopadhyay ,  Stephen P. Eisenberg ,  Gary L. Stetler ,  Robert C. Thompson

    IPC分类号： A61K38/00 ,  C07K14/81 ,  C12N15/15 ,  C12N15/70 ,  A61K37/64 ,  C12N15/11 ,  C12N15/79

    CPC分类号： C07K14/811 ,  C12N15/70 ,  A61K38/00 ,  C07K2319/02

    摘要： A synthetic DNA sequence and its genetic equivalents are disclosed which sequences are capable, when used in a recombinant DNA method, of directing production of a serine protease inibitor protein. Recombinant DNA methods for the production of serine protease inhibitor proteins are also disclosed. These methods incorporate either the synthetic DNA sequence of the present invention or natural DNA sequences isolated from human cDNA or genomic libraries.In addition, a single polypeptide chain protein is disclosed which is capable of inhibiting chymotrypsin and elastase but not trypsin. In one embodiment, this protein is a shortened from (single domain) of the protein produced by the method described herein.

    摘要翻译： 公开了合成DNA序列及其遗传等同物，当在重组DNA方法中使用时，能够引导产生丝氨酸蛋白酶内含蛋白的序列。 还公开了用于产生丝氨酸蛋白酶抑制蛋白的重组DNA方法。 这些方法包括本发明的合成DNA序列或从人cDNA或基因组文库分离的天然DNA序列。 此外，公开了能够抑制糜蛋白酶和弹性蛋白酶而不是胰蛋白酶的单个多肽链蛋白。 在一个实施方案中，该蛋白质是从通过本文所述的方法产生的蛋白质的（单结构域）缩短。