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    NON-AQUEOUS SYNTHESIS OF POLYSACCHARIDE-PROTEIN CONJUGATES FOR VACCINES
    45.
    发明申请
    NON-AQUEOUS SYNTHESIS OF POLYSACCHARIDE-PROTEIN CONJUGATES FOR VACCINES 审中-公开
    用于疫苗的多糖 - 蛋白质结合蛋白的非水性合成

    公开(公告)号：US20120077965A1

    公开(公告)日：2012-03-29

    申请号：US13240588

    申请日：2011-09-22

    Applicant: Harvey Kaplan

    Inventor： Harvey Kaplan

    IPC: C07K14/34 ,  C07K14/33

    CPC classification number: C07K14/00 ,  A61K38/16 ,  A61K38/164 ,  A61K39/00 ,  A61K47/646 ,  A61K2039/555 ,  A61K2039/55511 ,  A61K2039/55583 ,  A61K2039/60 ,  A61K2039/6031 ,  A61K2039/6087 ,  C07K14/195 ,  C07K14/33 ,  C07K14/34

    Abstract: The invention is a novel chemical coupling methodology for the synthesis of a stable polysaccharide-protein conjugates as the immunogenic component for vaccines. A covalent bond is formed between polysaccharide and protein in the dry state in the absence of water and oxygen. A polysaccharide antigen is covalently linked to the protein by activating the polysaccharide with periodate to introduce aldehyde groups into the polysaccharide, lyophilizing an aqueous mixture of a protein and activated polysaccharide, sealing the dry lyophilized mixture in a vessel under vacuum or inert gas and then incubating the sealed vessel at an elevated temperature.

    Abstract translation: 本发明是用于合成稳定的多糖 - 蛋白质缀合物作为疫苗的免疫原性成分的新型化学偶联方法。 在没有水和氧的情况下，在干燥状态下，在多糖和蛋白质之间形成共价键。 多糖抗原与蛋白质共价连接，通过用高碘酸盐活化多糖以将醛基引入多糖中，冻干蛋白质和活化多糖的含水混合物，将干燥的冻干混合物在真空或惰性气体中密封在容器中，然后孵育 在高温下密封容器。

    Protein fragment complementation assays for the detection of biological or drug interactions
    47.
    发明申请
    Protein fragment complementation assays for the detection of biological or drug interactions 有权
    用于检测生物或药物相互作用的蛋白质片段互补测定法

    公开(公告)号：US20110287950A1

    公开(公告)日：2011-11-24

    申请号：US13137257

    申请日：2011-08-01

    Applicant: Stephen William Watson Michnick ,  Joelle Nina Polletier ,  Ingrid Remy

    Inventor： Stephen William Watson Michnick ,  Joelle Nina Polletier ,  Ingrid Remy

    IPC: C40B30/00 ,  C07K14/34 ,  C07K14/435 ,  C12N9/96 ,  C12Q1/66 ,  C12Q1/48 ,  C12Q1/34 ,  C12Q1/26 ,  C07K14/21 ,  C07K14/00

    CPC classification number: G01N33/6845 ,  A01K67/0271 ,  A01K2267/0331 ,  A01K2267/0393 ,  C07K14/43595 ,  C07K2319/00 ,  C12N15/1055 ,  G01N33/5008 ,  G01N33/542 ,  G01N33/573 ,  G01N33/58 ,  G01N33/581 ,  G01N33/68 ,  G01N33/6803 ,  Y02A90/26

    Abstract: We describe a strategy for designing and implementing protein-fragment complementation assays (PCAs) to detect biomolecular interactions in vivo and in vitro. The design, implementation and broad applications of this strategy are illustrated with a large number of enzymes with particular detail provided for the example of murine dihydrofolate reductase (DHFR). Fusion peptides consisting of N- and C-terminal fragments of murine DHFR fused to GCN4 leucine zipper sequences were coexpressed in Escherichia coli grown in minimal medium, where the endogenous DHFR activity was inhibited with trimethoprim. Coexpression of the complementary fusion products restored colony formation. Survival only occurred when both DHFR fragments were present and contained leucine-zipper forming sequences, demonstrating that reconstitution of enzyme activity requires assistance of leucine zipper formation. DHFR fragment-interface point mutants of increasing severity (Ile to Val, Ala and Gly) resulted in a sequential increase in E. coli doubling times illustrating the successful DHFR fragment reassembly rather than non-specific interactions between fragments. This assay could be used to study equilibrium and kinetic aspects of molecular interactions including protein-protein, protein-DNA, protein-RNA, protein-carbohydrate and protein-small molecule interactions, for screening cDNA libraries for binding of a target protein with unknown proteins or libraries of small organic molecules for biological activity. The selection and design criteria applied here is developed for numerous examples of clonal selection, colorometric, fluorometric and other assays based on enzymes whose products can be measured. The development of such assay systems is shown to be simple, and provides for a diverse set of protein fragment complementation applications.

    Abstract translation: 我们描述了设计和实施蛋白质片段互补测定（PCAs）以检测体内和体外生物分子相互作用的策略。 该策略的设计，实现和广泛应用用大量酶进行了说明，具体细节提供了鼠二氢叶酸还原酶（DHFR）的例子。 由融合到GCN4亮氨酸拉链序列的鼠DHFR的N-和C-末端片段组成的融合肽在基本培养基中生长的大肠杆菌中共表达，其中内源性DHFR活性被甲氧苄啶抑制。 互补融合产物的共表达恢复了菌落形成。 存活只发生在两个DHFR片段存在并含有亮氨酸 - 拉链形成序列时，表明重组酶活性需要辅助亮氨酸拉链形成。 增加严重程度（Ile至Val，Ala和Gly）的DHFR片段 - 接口点突变体导致大肠杆菌倍增时间的顺序增加，说明成功的DHFR片段重组而不是片段之间的非特异性相互作用。 该测定可用于研究包括蛋白质蛋白质，蛋白质-DNA，蛋白质-RNA，蛋白质 - 碳水化合物和蛋白质 - 小分子相互作用在内的分子相互作用的平衡和动力学方面，用于筛选靶向蛋白质与未知蛋白质结合的cDNA文库 或用于生物活性的小有机分子的文库。 这里应用的选择和设计标准是针对克隆选择，色度，荧光测定和其他基于可以测量其产物的酶的测定的许多实例开发的。 这种测定系统的开发被证明是简单的，并且提供了多种蛋白质片段互补应用的集合。

    Corynebacterium glutamicum genes encoding proteins involved in membrane synthesis and membrane transport
    49.
    发明申请
    Corynebacterium glutamicum genes encoding proteins involved in membrane synthesis and membrane transport 审中-公开
    编码参与膜合成和膜转运的蛋白质的谷氨酸棒杆菌基因

    公开(公告)号：US20050244935A1

    公开(公告)日：2005-11-03

    申请号：US11082389

    申请日：2005-03-16

    Applicant: Markus Pompejus ,  Burkhard Kroger ,  Hartwig Schroder ,  Oskar Zelder ,  Gregor Haberhauer

    Inventor： Markus Pompejus ,  Burkhard Kroger ,  Hartwig Schroder ,  Oskar Zelder ,  Gregor Haberhauer

    IPC: C07H21/04 ,  C07K14/34 ,  C12N1/21 ,  C12N9/10 ,  C12P13/04

    CPC classification number: C07K14/34

    Abstract: Isolated nucleic acid molecules, designated MCT nucleic acid molecules, which encode novel MCT proteins from Corynebacterium glutamicum are described. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing MCT nucleic acid molecules, and host cells into which the expression vectors have been introduced. The invention still further provides isolated MCT proteins, mutated MCT proteins, fusion proteins, antigenic peptides and methods for the improvement of production of a desired compound from C. glutamicum based on genetic engineering of MCT genes in this organism.

    Abstract translation: 描述了编码来自谷氨酸棒杆菌的新型MCT蛋白质的分离的核酸分子，命名为MCT核酸分子。 本发明还提供反义核酸分子，含有MCT核酸分子的重组表达载体，以及引入了表达载体的宿主细胞。 本发明还进一步提供了分离的MCT蛋白，突变的MCT蛋白，融合蛋白，抗原肽和基于该生物体中MCT基因的遗传工程改进从谷氨酸棒球菌生产所需化合物的方法。

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