Mammary gland tissue-specific expression system using β-casein promoter site of Korean native goat
    1.
    发明授权
    Mammary gland tissue-specific expression system using β-casein promoter site of Korean native goat 失效
    乳腺组织特异性表达系统使用韩国本地山羊的β-酪蛋白启动子位点

    公开(公告)号:US07053262B2

    公开(公告)日:2006-05-30

    申请号:US10347718

    申请日:2003-01-17

    IPC分类号: C12P21/00 C12N15/63

    摘要: The mammary gland-specific expression systems developed by the present inventors, named pGbc, pGbc_L and pGbc_S were deposited under the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purpose of Patent Procedure in the Korean Collection for Type Cultures (KCTC), Korean Research Institute of Bioscience and Biotechnology at 52, Oun-dong, Yusong-Ku, Taejon 305–333, Republic of Korea, on Aug. 17, 1998, and the accession deposit Nos. KCTC 0515BP, 0514BP and 0513BP were issued, respectively. All restructions on the availability to the public of the deposited materials will be irrevocably removed upon the granting of a patent. The systems of the present invention make it possible that desired proteins are produced by expression in mammary gland tissue-derived animal cells or through the milk secreted from the transgenic animals with the expression systems, thereby solving the above-mentioned problems, that is, the activity sustenance, production cost, and isolation and purification of the desired proteins.

    摘要翻译: 由本发明人开发的命名为pGbc,pGbc_L和pGbc_S的本发明人的乳腺特异性表达系统根据布达佩斯条约保藏在国际识别为韩国专利文献收集专利程序中的微生物保藏(KCTC) 1998年8月17日,大韩民国Taejon 305-333 Ousong Ku,东京生物科技生物技术研究所52号,KCTC 0515BP,0514BP和0513BP登记号, 分别。 所有关于向公众提供存放资料的调整将在授予专利后不可撤销地删除。 本发明的系统使得可能通过在乳腺组织来源的动物细胞中表达所产生的蛋白质,或通过用表达系统从转基因动物分泌的乳中产生所需的蛋白质,由此解决上述问题,即, 活性维持,生产成本,以及所需蛋白质的分离和纯化。

    Cattle beta-casein gene targeting vector using homologous recombination
    3.
    发明授权
    Cattle beta-casein gene targeting vector using homologous recombination 有权
    使用同源重组的牛β-酪蛋白基因靶向载体

    公开(公告)号:US08030538B2

    公开(公告)日:2011-10-04

    申请号:US10574747

    申请日:2005-11-18

    摘要: The present invention relates to a bovine beta-casein gene targeting vector comprising (1) a first region having a length of 5 to 12 kb which is homologous to the promoter and its flanking nucleic acid sequences of bovine beta-casein gene, and comprising exon 1, intron 1, and exon 2 of bovine beta-casein gene; (2) a region for cloning a nucleic acid coding for desired proteins; (3) a region for coding a positive selection marker; (4) a second region having a length of 2.8 to 3.5 kb which is homologous to the nucleic acid sequences of bovine beta-casein gene, and comprising exon 5, 6, 7 and 8, and intron 5, 6 and 7 of bovine beta-casein gene; wherein the nucleic acid segment corresponding to the first region is located upstream to the nucleic acid segment corresponding to the second region in the 5′-3′ arrangement of beta-casein gene. The present invention also relates to method producing the transgenic cattle which is bovine beta-casein gene-targeted with a gene coding a desired protein using the said vector and obtaining a large scale of a desired protein from the milk of the said transgenic cattle.

    摘要翻译: 本发明涉及一种牛β-酪蛋白基因靶向载体,其包含(1)长度为5至12kb的第一区,其与牛β-酪蛋白基因的启动子及其侧翼核酸序列同源且包含外显子 1,牛β酪蛋白基因的内含子1和外显子2; (2)克隆编码所需蛋白质的核酸的区域; (3)用于编码正选择标记的区域; (4)具有与牛β-酪蛋白基因的核酸序列同源的2.8至3.5kb长度的第二区域,并且包含外周血5,6,7和8以及牛β内含子5,6和7 -casein基因; 其中对应于所述第一区域的核酸区段位于对应于β-酪蛋白基因的5'-3'排列中的第二区域的核酸区段的上游。 本发明还涉及生产转基因牛的方法,该转基因牛是使用所述载体用编码所需蛋白质的基因靶向的牛β-酪蛋白基因,并从所述转基因牛的奶中获得大规模的所需蛋白质。

    CATTLE BETA-CASEIN GENE TARGETING VECTOR USING HOMOLOGOUS RECOMBINATION
    4.
    发明申请
    CATTLE BETA-CASEIN GENE TARGETING VECTOR USING HOMOLOGOUS RECOMBINATION 有权
    CATTLE BETA-CASEIN基因导向矢量使用同源重建

    公开(公告)号:US20090013419A1

    公开(公告)日:2009-01-08

    申请号:US10574747

    申请日:2005-11-18

    摘要: The present invention relates to a bovine beta-casein gene targeting vector comprising (1) a first region having a length of 5 to 12 kb which is homologous to the promoter and its flanking nucleic acid sequences of bovine beta-casein gene, and comprising exon 1, intron 1, and exon 2 of bovine beta-casein gene; (2) a region for cloning a nucleic acid coding for desired proteins; (3) a region for coding a positive selection marker; (4) a second region having a length of 2.8 to 3.5 kb which is homologous to the nucleic acid sequences of bovine beta-casein gene, and comprising exon 5, 6, 7 and 8, and intron 5, 6 and 7 of bovine beta-casein gene; wherein the nucleic acid segment corresponding to the first region is located upstream to the nucleic acid segment corresponding to the second region in the 5′-3′ arrangement of beta-casein gene. The present invention also relates to method producing the transgenic cattle which is bovine beta-casein gene-targeted with a gene coding a desired protein using the said vector and obtaining a large scale of a desired protein from the milk of the said transgenic cattle.

    摘要翻译: 本发明涉及一种牛β-酪蛋白基因靶向载体,其包含(1)长度为5至12kb的第一区,其与牛β-酪蛋白基因的启动子及其侧翼核酸序列同源且包含外显子 1,牛β酪蛋白基因的内含子1和外显子2; (2)克隆编码所需蛋白质的核酸的区域; (3)用于编码正选择标记的区域; (4)具有与牛β-酪蛋白基因的核酸序列同源的2.8至3.5kb长度的第二区域,并且包含外周血5,6,7和8以及牛β内含子5,6和7 -casein基因; 其中对应于所述第一区域的核酸区段位于对应于β-酪蛋白基因的5'-3'排列中的第二区域的核酸区段的上游。 本发明还涉及生产转基因牛的方法,该转基因牛是使用所述载体用编码所需蛋白质的基因靶向的牛β-酪蛋白基因,并从所述转基因牛的乳中获得大规模的所需蛋白质。

    Mammary gland tissue-specific expression system using &bgr;-casein promoter site of Korean native goat
    5.
    发明授权
    Mammary gland tissue-specific expression system using &bgr;-casein promoter site of Korean native goat 失效
    乳腺组织特异性表达系统使用韩国本地山羊的β-酪蛋白启动子位点

    公开(公告)号:US06635474B1

    公开(公告)日:2003-10-21

    申请号:US09582490

    申请日:2000-11-14

    IPC分类号: C12N1563

    摘要: There are disclosed mammary gland tissue-specific expression systems using the promoter site for the &bgr;-casein gene of Korean native goats, by use of which physiological activating substances can be produced. In each of the expression systems, that is, novel plasmids pGbc, pGbc_L and pGbc_S (deposition Nos. KCTC 0515BP, 0514BP and 0513BP, respectively), a &bgr;-casein gene expression-regulating region, a physiological activating substance gene and a termination-regulating region are linked. Human granulocyte colony stimulating factor (hG-CSF) or human granulocyte macrophage colony stimulating factor (hGM-CSF) can be produced in HC11 cells, a mouse mammary gland tissue-derived cell line, and in the milk secreted from the transgenic mice by use of a hG-CSF or hGM-CSF gene-carrying pGbc, pGbc_L or pGbc_S in transfection into cell and microinjection to mouse. The proteins are those which experience the posttranslational modification and maintain their normal activity in the human body. The expression systems make it possible to easily produce the proteins at a great amount, to scale up protein production to the extent of industrialization, and to isolate and purify the desired protein with ease and safety.

    摘要翻译: 已经公开了使用韩国本地山羊的β-酪蛋白基因的启动子位点的乳腺组织特异性表达系统,通过使用可以产生生理活化物质。 在每个表达系统中,即新型质粒pGbc,pGbc_L和pGbc_S(分别为沉积编号KCTC 0515BP,0514BP和0513BP),β-酪蛋白基因表达调节区,生理活化物质基因和终止 - 调节区域相连。 人类粒细胞集落刺激因子(hG-CSF)或人粒细胞巨噬细胞集落刺激因子(hGM-CSF)可以在HC11细胞,小鼠乳腺组织来源的细胞系和转基因小鼠分泌的乳中产生 的hG-CSF或携带hGM-CSF基因的pGbc,pGbc_L或pGbc_S转染入细胞和显微注射到小鼠。 蛋白质是经历翻译后修饰并维持其在人体中的正常活性的蛋白质。 表达系统使得可以容易地大量生产蛋白质,将蛋白质生产扩大到工业化程度,并且容易且安全地分离和纯化所需的蛋白质。