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    Process for sexing cow embryos
    13.
    发明授权
    Process for sexing cow embryos 失效
    性交奶牛胚胎的过程

    公开(公告)号：US5876942A

    公开(公告)日：1999-03-02

    申请号：US899811

    申请日：1997-07-24

    Applicant: Winston Teng-Kuei Cheng ,  Chuan-Mu Chen ,  Che-Lin Hu ,  Chih-Hua Wang ,  Kong-Bung Choo

    Inventor： Winston Teng-Kuei Cheng ,  Chuan-Mu Chen ,  Che-Lin Hu ,  Chih-Hua Wang ,  Kong-Bung Choo

    IPC: C12Q1/68 ,  C07H21/02 ,  C07H21/04

    CPC classification number: C12Q1/6879

    Abstract: A rapid, highly reproducible and sensitive technique has been successfully developed for sexing the cow embryos, by method of polymerase chain reaction (PCR) against the amelogenin (bAML) genes located on both X- and Y-chromosomes of the Holstein dairy cattle. Results from DNA sequence analysis showed that there was only 45% homology between the intron 5 of AMLX and AMLY genes. Based on these sequences a pair of sex-specific primers, pbAML5XY(+) and pbAML5XY (-),were designed allowing to amplify a single fragment of 476-bp from the female cattle and two fragments of 476-bp and 341-bp from the male ones, respectively. The most important feature is that the precise sensitivity of sex-determination was confirmed to be reached as minimum template as trace amount of genomic DNA content in either a single lymphocyte or a single blastomere isolated from cow embryo at day-6 to day-7. Moreover, neither those of complicated procedures for purifying the DNA prior the PCR nor any extra pair of primers for serving as internal control is thought to be essential and the sex-determination of over hundred embryos can be completed at once within 4hrs.

    Abstract translation: 通过聚合酶链反应（PCR）对位于荷斯坦奶牛的X射线和Y染色体上的牙釉蛋白（bAML）基因的方法，已经成功开发了快速，高度重现和敏感的技术，用于性别雌性胚胎。 DNA序列分析结果表明，AMLX的内含子5与AMLY基因之间只有45％的同源性。 基于这些序列，设计了一对性别特异性引物pbAML5XY（+）和pbAML5XY（ - ），从而扩增了来自雌性牛的476-bp的单个片段和476bp和341bp的两个片段 男性分别。 最重要的特征是确定了在第6天至第7天从单纯淋巴细胞或从胚胎中分离的单个卵裂球中的最小模板作为最小模板作为痕量的基因组DNA含量。 此外，在PCR前纯化DNA的复杂程序和用作内部对照的任何额外的引物对都不被认为是至关重要的，并且可以在4小时内立即完成100多个胚胎的性别测定。

    Method for expressing multiple recombinant proteins in milk of transgenic non-human mammals
    16.
    发明申请
    Method for expressing multiple recombinant proteins in milk of transgenic non-human mammals 有权
    用于在转基因非人哺乳动物的乳中表达多种重组蛋白的方法

    公开(公告)号：US20050114911A1

    公开(公告)日：2005-05-26

    申请号：US10790720

    申请日：2004-03-03

    Applicant: Shinn-Chih Wu ,  Teng-Kuei Cheng ,  Chuan-Mu Chen ,  Shau-Ping Lin ,  Chon-Ho Yen ,  Ping-Cheng Yang

    Inventor： Shinn-Chih Wu ,  Teng-Kuei Cheng ,  Chuan-Mu Chen ,  Shau-Ping Lin ,  Chon-Ho Yen ,  Ping-Cheng Yang

    IPC: A01K67/027

    CPC classification number: A01K67/0275 ,  A01K2217/05 ,  A01K2227/108 ,  A01K2267/01

    Abstract: The present invention discloses a method for expressing multiple recombinant proteins in transgenic non-human mammalian milk, characterized in which human clotting factor IX gene and porcine lactoferrin gene are transferred into the mammal by gene injection and embryonic implantation to obtain expression in the milk of transgenic mammal and its filial generation. The method of this invention can maintain the stable expression of multiple recombinant proteins in the transgenic mammal during lactation and stable expression amount proximate to that of the first generation in the offsprings of the transgenic non-human mammal.

    Abstract translation: 本发明公开了一种在转基因非人哺乳动物乳中表达多种重组蛋白的方法，其特征在于通过基因注射和胚胎移植将人凝血因子IX基因和猪乳铁蛋白基因转移到哺乳动物中，以获得转基因乳中的表达 哺乳动物及其子代。 本发明的方法可以在哺乳期间保持转基因哺乳动物中多种重组蛋白的稳定表达，并且在转基因非人哺乳动物的后代中可以保持接近于第一代的稳定表达量。

    Novel milk-fermented product and use thereof
    17.
    发明申请
    Novel milk-fermented product and use thereof 审中-公开
    新型乳发酵产品及其用途

    公开(公告)号：US20140120173A1

    公开(公告)日：2014-05-01

    申请号：US13661080

    申请日：2012-10-26

    Applicant: Chuan-Mu Chen ,  Hsiao-Ling Chen

    Inventor： Chuan-Mu Chen ,  Hsiao-Ling Chen

    IPC: C07K2/00 ,  A23C9/12 ,  A61K35/20

    CPC classification number: C07K2/00 ,  A23C9/127 ,  A61K35/20 ,  A61K38/018

    Abstract: The present invention discloses a novel milk-fermented product comprising a group of peptides. The peptides are composed from no. 1 to no. 7 of the figure that is selected from the group consisting of FIG. 11 to FIG. 18. And the novel milk-fermented product is used for suppressing the reduction of bone mineral density. Therefore, the novel milk-fermented product could be a component of foods, nutrient supplement or medicine for treating or preventing osteoporosis.

    Abstract translation: 本发明公开了一种包含一组肽的新型乳发酵产品。 肽由no组成。 1到否 该图中的图7选自图1所示的组。 图11〜 而新型乳发酵产品用于抑制骨矿物质密度的降低。 因此，新型乳发酵产品可以作为食品，营养补充剂或治疗或预防骨质疏松症的药物的成分。

    Transgenic mammal secreting B-domain deleted human FVII in its milk
    18.
    发明授权
    Transgenic mammal secreting B-domain deleted human FVII in its milk 有权
    转基因哺乳动物在其牛奶中分泌B结构域缺失的人FVII

    公开(公告)号：US07667089B2

    公开(公告)日：2010-02-23

    申请号：US10820777

    申请日：2004-04-09

    Applicant: Winston T. K. Cheng ,  Chuan-Mu Chen ,  Shwu-Wha Lin ,  Chih-Hong Wang ,  Chih-Jen Lin ,  Shinn-Chih Wu

    Inventor： Winston T. K. Cheng ,  Chuan-Mu Chen ,  Shwu-Wha Lin ,  Chih-Hong Wang ,  Chih-Jen Lin ,  Shinn-Chih Wu

    IPC: C12P21/00 ,  A01K67/027 ,  C12N15/00

    CPC classification number: C12N15/8509 ,  A01K67/0275 ,  A01K2217/05 ,  A01K2267/01 ,  C07K14/755 ,  C12N2830/008

    Abstract: Hemophilia A is one of the major inherited bleeding disorders caused by a deficiency or abnormality in coagulation factor VIII (FVIII). Hemophiliacs have been treated with whole plasma or purified FVIII concentrates. The risk of transmitting blood-borne viruses and the cost of highly purified FVIII are the major factors that restrict prophylaxis in hemophilia therapy. One of the challenges created by the biotechnology revolution is the development of methods for the economical production of highly purified proteins in large scales. The present invention provides improved mammary expression cassettes useful for the expression of genes at high levels in the milk of transgenic animals. In particular, the present invention provides recombinant signal peptide sequences derived from a-lactalbumin and aS1-casein milk genes suitable for leading protein secretion in the mammary gland. These gene cassettes are capable of delivering different transgenic constructs which result in the production of full-length or B domain-deleted therapeutic levels of biologically active human FVIII in the transgenic animals in vivo. Within the scope of the invention are also method for producing the transgenic non-human mammal, such as mouse, rat, rabbit, goat, sheep, pig and bovine species, capable of expressing human FVIII, and methods of making milk and methods of identifying protein from the transgenic milk.

    Abstract translation: 血友病A是凝血因子VIII（FVIII）缺乏或异常引起的主要遗传性出血性疾病之一。 已经用全血浆或纯化的FVIII浓缩物治疗血友病患者。 传播血源性病毒的风险和高度纯化的FVIII的成本是限制血友病治疗预防的主要因素。 生物技术革命带来的挑战之一是开发大规模经济生产高纯度蛋白质的方法。 本发明提供了可用于在转基因动物的牛奶中高水平表达基因的改进的乳腺表达盒。 特别地，本发明提供了衍生自α-乳白蛋白的重组信号肽序列和适合于在乳腺中引导蛋白质分泌的α1酪蛋白乳基因。 这些基因盒能够递送不同的转基因构建体，其导致在转基因动物体内产生全长或B结构域缺失的生物活性人类FVIII的治疗水平。 在本发明的范围内，也是用于生产能够表达人FVIII的转基因非人哺乳动物如小鼠，大鼠，兔，山羊，绵羊，猪和牛物种的方法，以及制备牛奶的方法和鉴定方法 来自转基因乳的蛋白质。

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