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    Method and kit for expressing protein under regulation of the expression from repeated sequence formed by gene amplification, and transformant
    51.
    发明授权
    Method and kit for expressing protein under regulation of the expression from repeated sequence formed by gene amplification, and transformant 有权
    用于通过基因扩增形成的重复序列表达调控表达蛋白质的方法和试剂盒，以及转化体

    公开(公告)号：US08771983B2

    公开(公告)日：2014-07-08

    申请号：US11889512

    申请日：2007-08-14

    Applicant: Noriaki Shimizu

    Inventor： Noriaki Shimizu

    IPC: C12N15/85 ,  C12N15/87 ,  C12N15/90 ,  C12N15/113 ,  C12N5/16

    CPC classification number: C12N15/67 ,  C12N15/85 ,  C12N2810/85 ,  C12N2820/85 ,  C12P21/02

    Abstract: A method is disclosed for releasing the transcriptional regulation caused by a repeated sequence in a gene, a kit therefor and so on to thereby establish a system capable of producing a protein in a large amount. At least one embodiment of the method can be achieved by any one or more of the following methods: (a) in the amplification of a gene encoding a target protein, co-amplifying a polynucleotide of 10 kbp or more such as a λ-phage DNA or an insulator sequence; (b) selecting by culturing cells having undergone gene amplification in media containing a drug with a gradual increase in concentration; (c) elevating the promoter activity of inducing the expression of a gene encoding a target protein; (d) excising an amplified gene region from a chromosome with the use of Cre-LoxP System; (e) treating cells having undergone gene amplification with 5-aza-2′-deoxycytidine to thereby lower the methylation degree of DNA; and (f) selecting the mammalian cells having undergone gene amplification on double minute chromosomes.

    Abstract translation: 公开了一种用于释放由基因中的重复序列引起的转录调控的方法，其用于其的试剂盒等，从而建立能够大量产生蛋白质的系统。 该方法的至少一个实施方案可以通过以下任何一种或多种方法来实现：（a）扩增编码靶蛋白的基因，共扩增10kbp以上的多核苷酸，例如λ噬菌体 DNA或绝缘子序列; （b）通过培养在具有逐渐增加浓度的药物的培养基中培养经历基因扩增的细胞进行选择; （c）提高诱导编码靶蛋白的基因表达的启动子活性; （d）使用Cre-LoxP系统从染色体切除扩增的基因区域; （e）处理已经用5-氮杂-2'-脱氧胞苷进行基因扩增的细胞，从而降低DNA的甲基化程度; 和（f）选择在双分子染色体上经历基因扩增的哺乳动物细胞。

    FUSOGENIC PROPERTIES OF SAPOSIN C AND RELATED PROTEINS AND PEPTIDES FOR APPLICATION TO TRANSMEMBRANE DRUG DELIVERY SYSTEMS
    52.
    发明申请
    FUSOGENIC PROPERTIES OF SAPOSIN C AND RELATED PROTEINS AND PEPTIDES FOR APPLICATION TO TRANSMEMBRANE DRUG DELIVERY SYSTEMS 审中-公开
    SAPOSIN C和相关蛋白和肽在胰蛋白酶药物递送系统中的应用

    公开(公告)号：US20120020878A1

    公开(公告)日：2012-01-26

    申请号：US13127630

    申请日：2008-11-07

    Applicant: Xiaoyang Qi

    Inventor： Xiaoyang Qi

    IPC: A61K47/16 ,  A61K39/44 ,  A61K38/44 ,  A61K38/46 ,  G01N33/574 ,  A61P35/00 ,  A61K49/18 ,  A61K49/04 ,  A61K49/06 ,  G01N33/53 ,  A61K51/08 ,  A61K33/24

    CPC classification number: A61K9/1277 ,  A61K9/0019 ,  A61K9/1271 ,  C07K14/475 ,  C12N15/88 ,  C12N2810/85

    Abstract: The present invention comprises a method for delivering pharmaceutical and/or imaging agents within and/or through the dermal, mucosal and other cellular membranes, and across the blood-brain barrier, utilizing a fusogenic protein. The fusogenic protein is associated with a phospholipid membrane, such as a liposome. The liposome may include dioleoylphosphatidylserine, a negatively charged long-chain lipid. Alternatively, the liposome is comprised of a mixture of negatively charged long-chain lipids, neutral long-chain lipids, and neutral short-chain lipids. Preferred fusogenic proteins include saposin C and other proteins, polypeptides and peptide analogs derived from saposin C. The active agent contained within the liposome may comprise biomolecules and/or organic molecules. This technology can be used for both cosmetic and medicinal applications in which the objective is delivery of the active agent within and/or beneath biological membranes or across the blood-brain barrier and neuronal membranes.

    Abstract translation: 本发明包括利用融合蛋白质在真皮，粘膜和其它细胞膜内和/或通过真皮，粘膜和其它细胞膜以及横跨血脑屏障递送药物和/或成像剂的方法。 融合蛋白与磷脂膜如脂质体相关联。 脂质体可以包括二油酰磷脂酰丝氨酸，一种带负电荷的长链脂质。 或者，脂质体由带负电荷的长链脂质，中性长链脂质和中性短链脂质的混合物组成。 优选的融合蛋白包括滑膜蛋白C和衍生自皂角苷C的其它蛋白质，多肽和肽类似物。脂质体内包含的活性剂可以包含生物分子和/或有机分子。 该技术可用于化妆品和药物应用，其目的是将活性剂递送至生物膜内和/或生物膜内或穿过血脑屏障和神经元膜。

    NASAL-ADMINISTERED VACCINES USING MULTI-SCREENED NALT-TARGETING AND PHAGOCYTIC POLYPEPTIDE TRANSPORT SEQUENCES
    54.
    发明申请
    NASAL-ADMINISTERED VACCINES USING MULTI-SCREENED NALT-TARGETING AND PHAGOCYTIC POLYPEPTIDE TRANSPORT SEQUENCES 审中-公开
    使用多层筛选和药代动力学运输序列的NASAL-管理疫苗

    公开(公告)号：US20100278846A1

    公开(公告)日：2010-11-04

    申请号：US12636705

    申请日：2009-12-11

    Applicant: IAN A. FERGUSON

    Inventor： IAN A. FERGUSON

    IPC: A61K39/00 ,  A61P37/04

    CPC classification number: C12N7/00 ,  A61K39/00 ,  A61K2039/5256 ,  A61K2039/5258 ,  A61K2039/543 ,  C07K2319/735 ,  C12N2795/00042 ,  C12N2795/00051 ,  C12N2810/85

    Abstract: Multiple sequential screening tests have been performed on phage display libraries, and polypeptide sequences have been identified that potently drive both: (i) intake into mucosal immune cells, including NALT cells in the nose and throat; and, (ii) phagocytic intake and processing by antigen-presenting cells, such as macrophages. Such polypeptide sequences can be used as potent “target and deliver” components in vaccines that can be administered nasally, or to other mucous membranes. Such vaccines can be made very rapidly and in huge quantities, from bacteriophages that will also carry antigenic sequences in their coat proteins, or other immunoactive components. Alternately, such “target and deliver” polypeptides can be incorporated into vaccines derived from eukaryotic viruses or cellular pathogens. Enhancements also are disclosed, such as agents that can activate one or more types of toll-like receptors, to increase immunes responses and guide them in desired directions.

    Abstract translation: 已经对噬菌体展示文库进行了多次连续的筛选测试，并且已经鉴定出有效驱动两者的多肽序列：（i）摄入到粘膜免疫细胞，包括鼻和喉部的NALT细胞; 和（ii）由抗原呈递细胞如巨噬细胞吞噬摄入和加工。 这样的多肽序列可以用作可以被鼻腔或其它粘膜施用的疫苗中的有效的“靶和递送”成分。 这样的疫苗可以非常迅速地并且大量地从还会在其外壳蛋白或其它免疫活性成分中携带抗原序列的噬菌体制备。 或者，这样的“靶和递送”多肽可以并入来自真核病毒或细胞病原体的疫苗中。 还公开了增强剂，例如可以激活一种或多种类型的toll样受体的药剂，以增加免疫反应并将它们引导到期望的方向。

    Nucleic Acid Construct
    58.
    发明申请
    Nucleic Acid Construct 失效
    核酸构建体

    公开(公告)号：US20070281356A1

    公开(公告)日：2007-12-06

    申请号：US11632138

    申请日：2005-07-11

    Applicant: Seiji Shinkai ,  Takeshi Nagasaki ,  Takeshi Kawazu ,  Shinji Kakimoto

    Inventor： Seiji Shinkai ,  Takeshi Nagasaki ,  Takeshi Kawazu ,  Shinji Kakimoto

    IPC: C12N15/09 ,  C07H21/00 ,  C12N15/11 ,  C12N15/00

    CPC classification number: C12N15/88 ,  C07K2319/09 ,  C07K2319/23 ,  C12N15/87 ,  C12N2810/6072 ,  C12N2810/85

    Abstract: To provide a new technique by which efficient transfection is ensured in delivering a target gene into a cell, disclosed is a nucleic acid construct for nuclear import, which comprises a ternary complex consisting of a nucleic acid substance containing a gene to be delivered into the nucleus of a cell, an importin protein (for example, importin-β) capable of passing through the nuclear pore and involved in the nuclear transport, and a binding substance (for example, polyethyleneimine) bound to both of the nucleic acid substance and the importin protein. Nucleic acid transport from outside of a cell into the cell nucleus can be particularly promoted by administering the nucleic acid construct bound to a cell membrane receptor binding factor and/or a membrane fusing substance, or administering the nucleic acid construct encapsulated in a non-viral vector (for example, Sendai virus envelope) having cell membrane permeability and membrane fusing properties.

    Abstract translation: 为了提供一种新的技术，通过该技术确保将靶基因递送到细胞中的有效转染，公开了一种用于核进入的核酸构建体，其包含由含有待递送至细胞核的基因的核酸物质组成的三元复合物 的细胞，能够穿过核孔并参与核转运的importin蛋白（例如，importin-β）以及结合核酸物质和进口蛋白的结合物质（例如聚乙烯亚胺） 蛋白。 通过施用与细胞膜受体结合因子和/或膜融合物质结合的核酸构建体，或者将包封在非病毒体内的核酸构建体施用，可以特别地促进从细胞外进入细胞核的核酸 具有细胞膜通透性和膜融合特性的载体（例如仙台病毒包膜）。

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