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    Transgenic bovines and milk from transgenic bovines
    5.
    发明授权
    Transgenic bovines and milk from transgenic bovines 失效
    转基因牛的转基因牛奶和牛奶

    公开(公告)号：US6013857A

    公开(公告)日：2000-01-11

    申请号：US464167

    申请日：1995-06-05

    申请人： Herman A. Deboer ,  Rein Strijker ,  Herbert L. Heyneker ,  Gerard Platenburg ,  Sang He Lee ,  Frank Pieper ,  Paul J. A. Krimpenfort

    发明人： Herman A. Deboer ,  Rein Strijker ,  Herbert L. Heyneker ,  Gerard Platenburg ,  Sang He Lee ,  Frank Pieper ,  Paul J. A. Krimpenfort

    IPC分类号： A01K67/027 ,  A23C9/20 ,  C07K14/47 ,  C07K14/765 ,  C07K14/79 ,  C12N9/36 ,  C12N9/64 ,  C12N15/12 ,  C12N15/85 ,  A01K67/00 ,  A23C11/10 ,  C12P21/00

    CPC分类号： C12N15/8509 ,  A01K67/0275 ,  A01K67/0278 ,  A23C9/20 ,  C07K14/4732 ,  C07K14/765 ,  C07K14/79 ,  C12N15/85 ,  C12N9/2462 ,  C12N9/6464 ,  C12Y304/21069 ,  A01K2207/15 ,  A01K2217/00 ,  A01K2217/05 ,  A01K2217/072 ,  A01K2227/101 ,  A01K2227/105 ,  A01K2267/01 ,  C12N2800/30 ,  C12N2830/008 ,  C12N2830/15 ,  C12N2830/42 ,  C12N2830/85 ,  C12N2840/44

    摘要： Transgenes for producing recombinant polypeptides transgenic bovine species. A transgene for producing recombinant polypeptides in the milk of transgenic bovine species comprises at least one expression regulation sequence, a secretory DNA sequence encoding a secretory signal sequence which is functional in mammary secretory cells of the bovine species and a recombinant DNA sequence encoding the recombinant polypeptide. Also included are methods for producing transgenic bovine species. The method includes introducing the above transgene into an embryonal target cell of a bovine species, transplanting the transgenic embryonic target cell formed thereby into a recipient bovine parent and identifying at least one female offspring which is capable of producing the recombinant polypeptide in its milk. The invention also includes transgenic bovine species capable of producing recombinant polypeptides in transgenic milk as well as the milk from such transgenic bovine species and food formulations containing one or more recombinant polypeptide. Methods are also provided for producing transgenic non-human mammals having a desirable phenotype. The method comprises first methylating a transgene followed by introduction into fertilized oocytes. The oocytes are then cultured to form pre-implantation embryos. Thereafter, at least one cell is removed from each of the pre-implantation embryos and the DNA digested with a restriction endonuclease capable of cleaving the methylated transgene but incapable of cleaving the unmethylated form of the transgene. Those pre-implantation embryos which have integrated the transgene contain DNA which is resistant to cleavage by the restriction endonuclease in the region containing the transgene.

    摘要翻译： 用于产生重组多肽转基因牛物种的转基因。 用于在转基因牛物种的乳中产生重组多肽的转基因包括至少一个表达调控序列，编码在牛物种的乳腺分泌细胞中有功能的分泌信号序列的分泌DNA序列和编码重组多肽的重组DNA序列 。 还包括用于生产转基因牛物种的方法。 该方法包括将上述转基因引入牛种的胚胎靶细胞，将由此形成的转基因胚胎靶细胞移植到接受者的母本中，鉴定能够在其牛奶中产生重组多肽的至少一个雌性后代。 本发明还包括能够在转基因乳中产生重组多肽的转基因牛物种以及来自这种转基因牛物种的乳和含有一种或多种重组多肽的食物制剂。 还提供了用于产生具有所需表型的转基因非人哺乳动物的方法。 该方法首先将转基因甲基化，然后引入受精卵母细胞。 然后培养卵母细胞以形成植入前的胚胎。 此后，从每个植入前胚胎中移出至少一个细胞，并用能够切割甲基化的转基因而不能切割未甲基化形式的转基因的限制性内切核酸酶消化的DNA。 已经整合了转基因的那些植入前胚胎含有对限制性内切核酸酶在含有转基因的区域的切割具有抗性的DNA。

    Protease resistant urokinase
    7.
    发明授权
    Protease resistant urokinase 失效
    耐腐蚀性尿素酶

    公开(公告)号：US5219569A

    公开(公告)日：1993-06-15

    申请号：US766858

    申请日：1985-08-16

    申请人： Michael Blaber ,  Herbert L. Heyneker ,  Gordon A. Vehar

    发明人： Michael Blaber ,  Herbert L. Heyneker ,  Gordon A. Vehar

    IPC分类号： A61K38/00 ,  C07H21/04 ,  C12N20060101 ,  C12N5/10 ,  C12N9/00 ,  C12N9/72 ,  C12N15/00 ,  C12N15/10 ,  C12N15/58 ,  C12N15/67 ,  C12N15/70 ,  C12N15/71 ,  C12P21/02 ,  C12Q1/00

    CPC分类号： C12N9/6462 ,  C12N15/102 ,  C12N15/67 ,  C12N15/71 ,  C12N9/6459 ,  C12Y304/21069 ,  C12Y304/21073 ,  A61K38/00

    摘要： Novel single-chain protease resistant urokinase derivatives are provided. In particular, derivatives are provided wherein the Lys.sub.135 Lys.sub.136 and Arg.sub.156 to Lys.sub.158 sites are rendered less susceptible to proteolytic cleavage are provided by occluding the sites or by covalently modifying them. Preferred covalent modifications are amino acid sequence variants at the sites where proteolysis of urokinase occurs. These are optimally produced by synthesis of single-chain urokinase mutants in recombinant cell culture. The novel urokinase derivatives herein offer the advantage of avoiding the generation of substantial two-chain urokinase, either in vivo or during recombinant cell culture. However, the derivatives continue function to activate plasminogen in initiating blood clot lysis.

    摘要翻译： 提供了新的单链蛋白酶抗尿素激酶衍生物。 特别地，提供衍生物，其中Lys135Lys136和Arg156至Lys158位点对蛋白水解切割的敏感性较低是通过闭塞位点或通过共价修饰而提供的。 优选的共价修饰是在发生尿激酶蛋白水解的位点处的氨基酸序列变体。 这些通过在重组细胞培养物中合成单链尿激酶突变体来最佳地产生。 本文中的新型尿激酶衍生物提供了避免在体内或重组细胞培养过程中产生实质的双链尿激酶的优点。 然而，衍生物继续发挥功能以激活纤溶酶原引发血液凝块溶解。