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    Nondenatured HIV envelope antigens for detecting early HIV-specific antibodies
    118.
    发明授权
    Nondenatured HIV envelope antigens for detecting early HIV-specific antibodies 失效
    用于检测早期HIV特异性抗体的非变性HIV包膜抗原

    公开(公告)号：US6074646A

    公开(公告)日：2000-06-13

    申请号：US728122

    申请日：1996-10-09

    申请人： Miles W. Cloyd ,  Keith M. Ramsey

    发明人： Miles W. Cloyd ,  Keith M. Ramsey

    IPC分类号： G01N33/50 ,  A61P31/18 ,  C07K14/155 ,  C07K14/16 ,  C12N5/10 ,  C12N15/09 ,  C12P21/02 ,  C12R1/91 ,  G01N33/15 ,  G01N33/569 ,  A61K39/21 ,  C07K1/00

    CPC分类号： C07K14/005 ,  G01N33/56988 ,  C12N2740/16122

    摘要： Recombinant human immunodeficiency virus antigens capable of immunologically identifying the presence of early anti-HIV antibodies are stably expressed in a number of cell lines. These antigens have several clinically important applications as non-hazardous tools in the detection of human immunodeficiency virus exposure/infection, and in screening methods for HIV infection in idiopathic chronic lymphopenia (ICL). These techniques are improved over existing immunologically based and PCR based detection methods, as they provide for the detection of infection/exposure in samples determined to be negative by conventional forms of these types of assays that do not detect anti-HIV gp160 antibodies that react to conformational epitopes of HIV. The invention finds particular application in the detection of human immunodeficiency virus exposure/infection in infants. The earlier detection of the described methods is provided through the preserved immunoreactivity of the described recombinant conformationally intact human immunodeficiency virus that is capable of detecting a class of "early" anti-human immunodeficiency virus antibody not previously detectable by standard Western Blot or ELISA methods. The human immunodeficiency virus gp160 envelope antigen comprises one of the specific recombinant antigens examined with clinical human samples in these improved screening methods.

    摘要翻译： 能够免疫鉴定早期抗HIV抗体存在的重组人免疫缺陷病毒抗原在许多细胞系中稳定表达。 这些抗原作为检测人体免疫缺陷病毒暴露/感染的非危险工具以及特发性慢性淋巴细胞减少症（ICL）中艾滋病毒感染的筛选方法，具有几个临床上重要的应用。 这些技术比现有的基于免疫学和基于PCR的检测方法得到改进，因为它们提供了通过常规形式的这些类型的测定法检测到的阴性的感染/暴露的检测，其不检测抗HIV gp160抗体的反应 HIV的构象表位。 本发明特别适用于检测婴幼儿人体免疫缺陷病毒的暴露/感染。 所述方法的早期检测通过所述重组构象完整的人类免疫缺陷病毒的保护性免疫反应性提供，其能够检测以前不能通过标准Western印迹或ELISA方法检测的一类“早期”抗人免疫缺陷病毒抗体。 人类免疫缺陷病毒gp160包膜抗原包括在这些改进的筛选方法中用临床人类样品检测的特异性重组抗原之一。

    Ubiquitin expression system
    119.
    发明授权
    Ubiquitin expression system 失效
    泛素表达系统

    公开(公告)号：US6068994A

    公开(公告)日：2000-05-30

    申请号：US428278

    申请日：1995-04-25

    申请人： Philip J. Barr

    发明人： Philip J. Barr

    IPC分类号： C07K14/16 ,  C07K14/57 ,  C07K14/81 ,  C12N15/81 ,  C07K19/00 ,  C12M15/62

    CPC分类号： C07K14/005 ,  C07K14/57 ,  C07K14/8125 ,  C12N15/81 ,  C12N2740/16122 ,  C12N2740/16222

    摘要： The present invention provides an improved ubiquitin fusion system for gene expression in yeast systems which allows for the regulatable high level production of heterologous proteins having destabilizing amino terminal residues. The ubiquitin fusion proteins expressed in yeast are cleaved precisely in vivo by an endogenous ubiquitin-specific hydrolase to yield heterologous proteins such as human alpha-1-antitrypsin, human gamma-interferon and human immunodeficiency virus integrase protein, all of which initiate with destabilizing residues. An expression vector containing a synthetic gene for monomeric yeast ubiquitin was constructed and expressed under the control of a glucose regulatable yeast promoter. Inclusion of unique restriction sites at the 3'-end of the synthetic ubiquitin gene allows for precise in-frame insertion of heterologous genes. The system can be used to increase expression of poorly expressed proteins and to produce proteins having selective amino-terminal destabilizing residues.

    摘要翻译： 本发明提供了改进的用于酵母系统中基因表达的泛素融合系统，其允许可调节的高水平产生具有不稳定氨基末端残基的异源蛋白质。 在酵母中表达的泛素融合蛋白通过内源泛素特异性水解酶在体内精确地切割，得到异源蛋白质，例如人α-1-抗胰蛋白酶，人γ-干扰素和人免疫缺陷病毒整合酶蛋白，所有这些都起始于不稳定残基 。 构建含有单体酵母泛素合成基因的表达载体，并在葡萄糖可调节酵母启动子的控制下进行表达。 在合成泛素基因的3'末端包含独特的限制性位点允许异源基因的精确的帧内插入。 该系统可用于增加表达差的蛋白质的表达并产生具有选择性氨基末端去稳定化残基的蛋白质。