公开(公告)号：US08030538B2

公开(公告)日：2011-10-04

申请号：US10574747

申请日：2005-11-18

申请人： Yong-Mahn Han ,  Kyung-Kwang Lee ,  Mira Chang ,  Deog-Bon Koo

发明人： Yong-Mahn Han ,  Kyung-Kwang Lee ,  Mira Chang ,  Deog-Bon Koo

IPC分类号： C12P21/00 ,  A01K67/027 ,  C12N15/00 ,  C12N5/00

CPC分类号： C12N15/8771 ,  A01K67/0275 ,  A01K2217/05 ,  A01K2227/101 ,  A01K2267/01 ,  C12N15/8509 ,  C12N2830/008

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。

公开(公告)号：US08507275B2

公开(公告)日：2013-08-13

申请号：US12858747

申请日：2010-08-18

申请人： Yong-Mahn Han ,  Sang-Wook Park ,  Eun-Young Lee

发明人： Yong-Mahn Han ,  Sang-Wook Park ,  Eun-Young Lee

IPC分类号： C12N5/00 ,  C12N5/02 ,  C12N5/08 ,  A01N61/00 ,  A01N37/18

CPC分类号： C12N5/069 ,  C12N5/0647 ,  C12N5/0691 ,  C12N5/0692 ,  C12N2501/115 ,  C12N2501/155 ,  C12N2501/165 ,  C12N2501/999 ,  C12N2506/02 ,  C12N2506/28

摘要： The present invention relates to a composition for inducing embryonic stem cell differentiation comprising a MEK/ERK (mitogen-activated protein kinase kinase/extracellular regulated kinase) signal transduction inhibitor and BMP (bone morphogenetic protein), and a method for inducing differentiation of embryonic stem cells into mesodermal cells using the same. Further, the mesodermal cells obtained by the above method are able to differentiate into various mesenchymal tissue cells. In particular, the present invention relates to a method for inducing differentiation into hemangioblast by culturing the mesodermal cells obtained by the above method in the presence of VEGF (vascular endothelial cell growth factor) and bFGF (basic fibroblast growth factor). The differentiated hemangioblasts can be further differentiated into vascular endothelial cells, vascular smooth muscle cells, and hematopoietic stem cells under various culture conditions.

摘要翻译： 本发明涉及一种用于诱导胚胎干细胞分化的组合物，其包含MEK / ERK（丝裂原活化蛋白激酶激酶/细胞外调节激酶）信号转导抑制剂和BMP（骨形态发生蛋白）），以及诱导胚胎干细胞分化的方法 细胞使用相同的细胞进入中胚层细胞。 此外，通过上述方法获得的中胚层细胞能够分化成各种间充质组织细胞。 特别地，本发明涉及通过在VEGF（血管内皮细胞生长因子）和bFGF（碱性成纤维细胞生长因子）的存在下培养通过上述方法获得的中胚层细胞来诱导分化为成血管细胞成分的方法。 分化的成血管细胞可以在各种培养条件下进一步分化成血管内皮细胞，血管平滑肌细胞和造血干细胞。

公开(公告)号：US20110027886A1

公开(公告)日：2011-02-03

申请号：US12858747

申请日：2010-08-18

申请人： Yong-Mahn Han ,  Sang-Wook Park ,  Eun-Young Lee

发明人： Yong-Mahn Han ,  Sang-Wook Park ,  Eun-Young Lee

IPC分类号： C12N5/077 ,  C12N5/073

CPC分类号： C12N5/069 ,  C12N5/0647 ,  C12N5/0691 ,  C12N5/0692 ,  C12N2501/115 ,  C12N2501/155 ,  C12N2501/165 ,  C12N2501/999 ,  C12N2506/02 ,  C12N2506/28

摘要： The present invention relates to a composition for inducing embryonic stem cell differentiation comprising a MEK/ERK (mitogen-activated protein kinase kinase/extracellular regulated kinase) signal transduction inhibitor and BMP (bone morphogenetic protein), and a method for inducing differentiation of embryonic stem cells into mesodermal cells using the same. Further, the mesodermal cells obtained by the above method are able to differentiate into various mesenchymal tissue cells. In particular, the present invention relates to a method for inducing differentiation into hemangioblast by culturing the mesodermal cells obtained by the above method in the presence of VEGF (vascular endothelial cell growth factor) and bFGF (basic fibroblast growth factor). The differentiated hemangioblasts can be further differentiated into vascular endothelial cells, vascular smooth muscle cells, and hematopoietic stem cells under various culture conditions.

摘要翻译： 本发明涉及一种用于诱导胚胎干细胞分化的组合物，其包含MEK / ERK（丝裂原活化蛋白激酶激酶/细胞外调节激酶）信号转导抑制剂和BMP（骨形态发生蛋白）），以及诱导胚胎干细胞分化的方法 细胞使用相同的细胞进入中胚层细胞。 此外，通过上述方法获得的中胚层细胞能够分化成各种间充质组织细胞。 特别地，本发明涉及通过在VEGF（血管内皮细胞生长因子）和bFGF（碱性成纤维细胞生长因子）的存在下培养通过上述方法获得的中胚层细胞来诱导分化为成血管细胞成分的方法。 分化的成血管细胞可以在各种培养条件下进一步分化成血管内皮细胞，血管平滑肌细胞和造血干细胞。

公开(公告)号：US20090013419A1

公开(公告)日：2009-01-08

申请号：US10574747

申请日：2005-11-18

申请人： Yong-Mahn Han ,  Kyung-Kwang Lee ,  Mira Chang ,  Deog-Bon Koo

发明人： Yong-Mahn Han ,  Kyung-Kwang Lee ,  Mira Chang ,  Deog-Bon Koo

IPC分类号： A01K67/027 ,  C12N15/00 ,  C12N5/06 ,  C12N15/87

CPC分类号： C12N15/8771 ,  A01K67/0275 ,  A01K2217/05 ,  A01K2227/101 ,  A01K2267/01 ,  C12N15/8509 ,  C12N2830/008

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。