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1. 发明授权

US07435869B2 Transgenic nonhuman mammals producing fibrinogen in milk and methods of producing fibrin 失效
标题翻译：在牛奶中产生纤维蛋白原的转基因非人哺乳动物和产生纤维蛋白的方法
公开(公告)号：US07435869B2
公开(公告)日：2008-10-14
申请号：US11040117
申请日：2005-01-24
申请人： William H. Velander , William N. Drohan , Henryk Lubon , John L. Johnson
发明人： William H. Velander , William N. Drohan , Henryk Lubon , John L. Johnson
IPC分类号： C12P21/00 , A01K67/027 , A61K38/00
CPC分类号： C07K14/75 , A01K67/0278 , A01K2207/15 , A01K2217/00 , A01K2217/05 , A01K2227/105 , A01K2227/108 , A01K2267/01 , C12N15/8509
摘要： A transgenic, non-human mammalian animal is capable of expressing a heterologous gene for human or other recombinant physiologically functional fibrinogen holoprotein or individual subunit chain polypeptides thereof or a modified or fusion fibrinogen in mammary glands of the animals and secreting the expressed product into a body fluid. Methodology employing such a mammal yields recombinant physiologically functional fibrinogens, subunit chain polypeptides thereof, and modified or fusion fibrinogens.
摘要翻译：转基因非人哺乳动物能够在动物的乳腺中表达人或其他重组生理功能性纤维蛋白原蛋白或其单个亚单位链多肽或修饰或融合纤维蛋白原的异源基因，并将表达的产物分泌到体内流体。使用这种哺乳动物的方法产生重组生理功能性纤维蛋白原，其亚单位链多肽，以及修饰或融合的纤维蛋白原。

2. 发明授权

US5880327A Transgenic mammals expressing human coagulation factor VIII 失效
标题翻译：表达人凝血因子VIII的转基因哺乳动物
公开(公告)号：US5880327A
公开(公告)日：1999-03-09
申请号：US308518
申请日：1994-09-21
申请人： Henryk Lubon , William N. Drohan , William H. Velander
发明人： Henryk Lubon , William N. Drohan , William H. Velander
IPC分类号： A01K67/027 , C07K14/755 , C12N15/09 , C12N15/85 , C12P21/02 , C12N15/00 , C12N5/00
CPC分类号： C12N15/8509 , A01K67/0278 , C07K14/755 , A01K2207/15 , A01K2217/00 , A01K2217/05 , A01K2227/10 , A01K2227/105 , A01K2267/01
摘要： A non-human transgenic mammalian animal, as described above, contains an exogenous double stranded DNA sequence stably integrated into the genome of the animal, which comprises cis-acting regulatory units operably linked to a DNA sequence encoding human Factor VIII protein and a signal peptide, where the cis-acting regulatory units are active in mammary gland cells and the signal peptide is active in directing newly expressed Factor VIII into the milk of the animal. The promoter may be a milk protein promoter such as for whey acidic protein, casein, lactalbumin, or beta-lactoglobulin promoter. The transgenic mammals are preferably farm animals, for example, cows, goats, sheep, rabbits and pigs. Concurrent expression of a gene for human von Willebrand's Factor into milk may be used to stabilize newly-secreted Factor VIII.
摘要翻译：如上所述的非人转基因哺乳动物包含稳定整合到动物基因组中的外源双链DNA序列，其包含与编码人因子VIII蛋白的DNA序列可操作地连接的顺式作用调节单元和信号肽其中顺式作用调节单元在乳腺细胞中是活性的，并且信号肽在将新表达的因子VIII引导到动物的乳中是有活性的。启动子可以是乳蛋白质启动子，例如用于乳清酸性蛋白质，酪蛋白，乳清蛋白或β-乳球蛋白启动子。转基因哺乳动物优选为农场动物，例如牛，山羊，绵羊，兔和猪。将人血管性血友病因子的基因同时表达为牛奶可用于稳定新分泌的因子VIII。

3. 发明授权

US06344596B1 Expression of active human factor IX in mammary tissue and of milk non human transgenic mammals 失效
标题翻译：活性人因子IX在转基因非人哺乳动物的乳腺组织和乳中的表达
公开(公告)号：US06344596B1
公开(公告)日：2002-02-05
申请号：US09367087
申请日：1999-09-15
申请人： William H. Velander , William N. Drohan , Henryk Lubon , John L. Johnson
发明人： William H. Velander , William N. Drohan , Henryk Lubon , John L. Johnson
IPC分类号： A01K67027
CPC分类号： C12N15/8509 , A01K67/0275 , A01K67/0278 , A01K2207/15 , A01K2217/00 , A01K2217/05 , A01K2227/105 , A01K2227/108 , A01K2267/01 , A01K2267/03 , A61K38/36 , C12N9/644 , C12Y304/21022
摘要： Recombinant Factor IX characterized by a high percentage of active protein can be obtained in the milk of transgenic animals that incorporate chimeric DNA molecules according to the present invention. Transgenic animals of the present invention are produced by introducing into developing embryos DNA that encodes Factor IX, such that the foreign DNA is stably incorporated in the DNA of germ line cells of the mature animal. Particularly efficient expression was accomplished using a chimeric construct comprising a mammary gland specific promoter, Factor IX cDNA that lacked the complete or any portion of the 5′-untranslated and 3′-untranslated region, which is substituted with a 5′- and 3′-end of the mouse whey acidic protein gene. In vitro cell cultures of cells explanted from the transgenic mammal of the invention and methods of producing Factor IX from such said culture and methods of treating hemophilia B are also described.
摘要翻译：可以在掺入根据本发明的嵌合DNA分子的转基因动物的乳中获得以高百分比的活性蛋白为特征的重组因子IX。本发明的转基因动物通过引入编码因子IX的发育中的胚胎DNA而产生，使得外源DNA稳定地并入成熟动物的种系细胞的DNA中。使用含有乳腺特异性启动子的嵌合构建体，缺乏5'和非翻译区的完整或任何部分的因子IX cDNA完成了特别有效的表达，该5'和3'非翻译区用5'和3' -end的小鼠乳清酸性蛋白基因。还描述了从本发明的转基因哺乳动物移出的细胞的体外细胞培养物和从所述培养物生产因子IX的方法和治疗血友病B的方法。

4. 发明授权

US06984772B1 Transgenic non-human mammals producing fibrinogen in their milk 失效
标题翻译：在其牛奶中产生纤维蛋白原的转基因非人哺乳动物
公开(公告)号：US06984772B1
公开(公告)日：2006-01-10
申请号：US08443184
申请日：1995-05-17
申请人： William H. Velander , William N. Drohan , Henryk Lubon , John L. Johnson
发明人： William H. Velander , William N. Drohan , Henryk Lubon , John L. Johnson
IPC分类号： C12P21/00 , A01K67/00 , C12N15/00
CPC分类号： C07K14/75 , A01K67/0278 , A01K2207/15 , A01K2217/00 , A01K2217/05 , A01K2227/105 , A01K2227/108 , A01K2267/01 , C12N15/8509
摘要： A transgenic, non-human mammalian animal is capable of expressing a heterologous gene for human or other recombinant physiologically functional fibrinogen holoprotein or individual subunit chain polypeptides thereof or a modified or fusion fibrinogen in mammary glands of the animals and secreting the expressed product into a body fluid. Methodology employing such a mammal yields recombinant physiologically functional fibrinogens, subunit chain polypeptides thereof, and modified or fusion fibrinogens.
摘要翻译：转基因非人哺乳动物能够在动物的乳腺中表达人或其他重组生理功能性纤维蛋白原蛋白或其单个亚单位链多肽或修饰或融合纤维蛋白原的异源基因，并将表达的产物分泌到体内流体。使用这种哺乳动物的方法产生重组生理功能性纤维蛋白原，其亚单位链多肽，以及经修饰或融合的纤维蛋白原。

5. 发明授权

US06262336B1 Expression of a heterologous protein C in mammary tissue of transgenic animals using a long whey acidic protein promoter 失效
标题翻译：使用长乳清酸性蛋白启动子在转基因动物的乳腺组织中表达异源蛋白C
公开(公告)号：US06262336B1
公开(公告)日：2001-07-17
申请号：US09321831
申请日：1999-05-28
申请人： Henryk Lubon , William N. Drohan , Lothar Hennighausen , William H. Velander
发明人： Henryk Lubon , William N. Drohan , Lothar Hennighausen , William H. Velander
IPC分类号： A01K67027
CPC分类号： C12N15/8509 , A01K67/0275 , A01K2207/15 , A01K2217/00 , A01K2217/05 , A01K2227/10 , A01K2227/105 , A01K2227/108 , A01K2267/01 , A61K38/00 , C07K14/75 , C07K2319/00 , C07K2319/02 , C12N9/6464 , C12N2830/008 , C12Y304/21069
摘要： An isolated DNA sequence which regulates the expression of a heterologous gene composed of a mouse whey acidic protein promoter having a length of greater than about 2.4 kb extending upstream from the unique KpnI site in the mouse whey acidic protein gene is disclosed. Specifically a mouse whey acidic protein promoter of about 4.1-4.2 kb in length extending upstream from the unique KpnI site is preferred. This mouse whey acid protein promoter is operably linked to a DNA sequence encoding a heterologous polypeptide and used to prepare transgenic non-human mammals expressing the heterologous polypeptide in their milk. Particularly efficient expression of both cDNAs and genomic DNAs encoding heterologous polypeptides was obtained in transgenic non-human mammals using this promoter, known as the long whey acidic protein promoter.
摘要翻译：公开了一种分离的DNA序列，其调节由小鼠乳清酸性蛋白基因中唯一KpnI位点上游延伸的长度大于约2.4kb的小鼠乳清酸性蛋白启动子组成的异源基因的表达。具体地，优选从独特的KpnI位点上游延伸的长度为约4.1-4.2kb的小鼠乳清酸性蛋白启动子。该小鼠乳清蛋白启动子可操作地连接到编码异源多肽的DNA序列，并用于制备在其牛奶中表达异源多肽的转基因非人哺乳动物。使用称为长乳清酸性蛋白启动子的该启动子，在转基因非人哺乳动物中获得编码异源多肽的cDNA和基因组DNA的特别有效的表达。

6. 发明授权

US06255554B1 Transgenic non-human mammals expressing human coagulation factor VIII and von Willebrand factor 失效
标题翻译：表达人凝血因子VIII和von Willebrand因子的转基因非人哺乳动物
公开(公告)号：US06255554B1
公开(公告)日：2001-07-03
申请号：US09262017
申请日：1999-03-04
申请人： Henryk Lubon , William N. Drohan , William H. Velander
发明人： Henryk Lubon , William N. Drohan , William H. Velander
IPC分类号： A01K67027
CPC分类号： C12N15/8509 , A01K67/0278 , A01K2207/15 , A01K2217/00 , A01K2217/05 , A01K2227/10 , A01K2227/105 , A01K2267/01 , C07K14/755
摘要： A non-human transgenic mammalian animal, as described above, contains an exogenous double stranded DNA sequence stably integrated into the genome of the animal, which comprises cis-acting regulatory units operably linked to a DNA sequence encoding human Factor VIII protein and a signal peptide, where the cis-acting regulatory units are active in mammary gland cells and the signal peptide is active in directing newly expressed Factor VIII into the milk of the animal. The promoter may be a milk protein promoter such as for whey acidic protein, casein, lactalbumin, or beta-lactoglobulin promoter. The transgenic mammals are preferably farm animals, for example, cows, goats, sheep, rabbits and pigs. Concurrent expression of a gene for human von Willebrand's Factor into milk may be used to stabilize newly-secreted Factor VIII.
摘要翻译：如上所述的非人转基因哺乳动物包含稳定整合到动物基因组中的外源双链DNA序列，其包含与编码人因子VIII蛋白的DNA序列可操作地连接的顺式作用调节单元和信号肽其中顺式作用调节单元在乳腺细胞中是活性的，并且信号肽在将新表达的因子VIII引导到动物的乳中是有活性的。启动子可以是乳蛋白质启动子，例如用于乳清酸性蛋白质，酪蛋白，乳清蛋白或β-乳球蛋白启动子。转基因哺乳动物优选为农场动物，例如牛，山羊，绵羊，兔和猪。将人血管性血友病因子的基因同时表达为牛奶可用于稳定新分泌的因子VIII。

7. 发明申请

US20090221492A1 EXPRESSION OF ACTIVE HUMAN FACTOR IX IN MAMMARY TISSUE OF TRANSGENIC ANIMALS 审中-公开
标题翻译：活性人因子IX在转基因动物哺乳动物组织中的表达
公开(公告)号：US20090221492A1
公开(公告)日：2009-09-03
申请号：US12201673
申请日：2008-08-29
申请人： William H. Velander , William N. Drohan , Henryk Lubon , John L. Johnson , Mary Ann H. Johnson
发明人： William H. Velander , William N. Drohan , Henryk Lubon , John L. Johnson , Mary Ann H. Johnson
IPC分类号： A61K38/16 , A01K67/027 , C12P21/00 , C12N5/00 , C07K14/00
CPC分类号： C12N15/8509 , A01K67/0275 , A01K67/0278 , A01K2207/15 , A01K2217/00 , A01K2217/05 , A01K2227/105 , A01K2227/108 , A01K2267/01 , A01K2267/03 , A61K38/36 , C12N9/644 , C12Y304/21022
摘要： Recombinant Factor IX characterized by a high percentage of active protein can be obtained in the milk of transgenic animals that incorporate chimeric DNA molecules according to the present invention. Transgenic animals of the present invention are produced by introducing into developing embryos DNA that encodes Factor IX, such that the foreign DNA is stably incorporated in the DNA of germ line cells of the mature animal. Particularly efficient expression was accomplished using a chimeric construct comprising a mammary gland specific promoter, Factor IX cDNA that lacked the complete or any portion of the 5′-untranslated and 3′-untranslated region, which is substituted with a 5-′ and 3′-end of the mouse whey acidic protein gene. In vitro cell cultures of cells explanted from the transgenic mammal of the invention and methods of producing Factor IX from such said culture and methods of treating hemophilia B are also described.
摘要翻译：可以在掺入根据本发明的嵌合DNA分子的转基因动物的乳中获得以高百分比的活性蛋白为特征的重组因子IX。本发明的转基因动物通过引入编码因子IX的发育中的胚胎DNA而产生，使得外源DNA稳定地并入成熟动物的种系细胞的DNA中。使用包含乳腺特异性启动子的嵌合构建体，缺乏5'和3'非翻译区的完整或任何部分的因子IX cDNA，其被5'和3' -end的小鼠乳清酸性蛋白基因。还描述了从本发明的转基因哺乳动物移出的细胞的体外细胞培养物和从所述培养物生产因子IX的方法和治疗血友病B的方法。

8. 发明授权

US06518482B2 Transgenic non-human mammals expressing human coagulation factor VIII and von Willebrand factor 失效
标题翻译：表达人凝血因子VIII和von Willebrand因子的转基因非人哺乳动物
公开(公告)号：US06518482B2
公开(公告)日：2003-02-11
申请号：US09849406
申请日：2001-05-07
申请人： Henryk Lubon , William N. Drohan , William H. Velander
发明人： Henryk Lubon , William N. Drohan , William H. Velander
IPC分类号： A01K67027
CPC分类号： C07K14/705
摘要： A non-human transgenic mammalian animal, as described above, contains an exogenous double stranded DNA sequence stably integrated into the genome of the animal, which comprises cis-acting regulatory units operably linked to a DNA sequence encoding human Factor VIII protein and a signal peptide, where the cis-acting regulatory units are active in mammary gland cells and the signal peptide is active in directing newly expressed Factor VIII into the milk of the animal. The promoter may be a milk protein promoter such as for whey acidic protein, casein, lactalbumin, or beta-lactoglobulin promoter. The transgenic mammals are preferably farm animals, for example, cows, goats, sheep, rabbits and pigs. Concurrent expression of a gene for human von Willebrand's Factor into milk may be used to stabilize newly-secreted Factor VIII.
摘要翻译：如上所述的非人转基因哺乳动物包含稳定整合到动物基因组中的外源双链DNA序列，其包含与编码人因子VIII蛋白的DNA序列可操作地连接的顺式作用调节单元和信号肽其中顺式作用调节单元在乳腺细胞中是活性的，并且信号肽在将新表达的因子VIII引导到动物的乳中是有活性的。启动子可以是乳蛋白质启动子，例如用于乳清酸性蛋白质，酪蛋白，乳清蛋白或β-乳球蛋白启动子。转基因哺乳动物优选为农场动物，例如牛，山羊，绵羊，兔和猪。将人血管性血友病因子的基因同时表达为牛奶可用于稳定新分泌的因子VIII。

9. 发明授权

US5965789A Engineering protein posttranslational modification by PACE/furin in transgenic non-human mammals 失效
标题翻译：在转基因非人类哺乳动物中通过PACE /弗林蛋白酶进行工程蛋白翻译后修饰
公开(公告)号：US5965789A
公开(公告)日：1999-10-12
申请号：US434834
申请日：1995-05-04
申请人： Henryk Lubon , William N. Drohan , Rekha K. Paleyanda
发明人： Henryk Lubon , William N. Drohan , Rekha K. Paleyanda
IPC分类号： A61K38/00 , C07K14/75 , C12N9/64 , C12N15/64 , C12N15/67 , C12N15/82 , C12N15/85 , A01K67/00 , A01K67/027 , C12P21/04 , C12P21/06
CPC分类号： A01K67/0278 , C07K14/75 , C12N15/64 , C12N15/67 , C12N15/8216 , C12N15/8243 , C12N15/85 , C12N15/8509 , C12N9/6454 , C12N9/6464 , C12Y304/21069 , A01K2207/15 , A01K2217/00 , A01K2217/05 , A01K2227/10 , A01K2227/105 , A01K2267/01 , A61K38/00 , C07K2319/00 , C07K2319/02 , C12N2830/008 , C12N2830/85
摘要： The invention relates to transgenic non-human multicellular organisms that contain polynucleotides for expressing proteins that alter posttranslational modification. In particular, the invention provides multiply-transgenic animals in which a first transgene encodes a first protein, a second transgene encodes a second protein, and expression of the second protein affects the posttranlational modification of the first protein in cells of said organism. Expression in preferred embodiments is in specific cells and the modified protein is secreted into a bodily fluid. The invention provides related methods, proteins and products. An example provides transgenic animals that express human Protein C and the processing protease PACE/furin in mammary glands and secrete both proteins into milk.
摘要翻译：本发明涉及包含用于表达改变翻译后修饰的蛋白质的多核苷酸的转基因非人多细胞生物体。特别地，本发明提供了多转基因动物，其中第一转基因编码第一蛋白质，第二转基因编码第二蛋白质，第二蛋白质的表达影响所述生物体细胞中第一蛋白质的后期修饰。优选实施方案中的表达在特定细胞中，并且所述修饰的蛋白质被分泌到体液中。本发明提供了相关方法，蛋白质和产物。一个实例提供表达人蛋白C的转基因动物和乳腺中加工蛋白酶PACE /弗林蛋白酶，并将两种蛋白质分泌到牛奶中。

10. 发明授权

US5831141A Expression of a heterologous polypeptide in mammary tissue of transgenic non-human mammals using a long whey acidic protein promoter 失效
标题翻译：使用长乳清酸性蛋白启动子在转基因非人哺乳动物的乳腺组织中表达异源多肽
公开(公告)号：US5831141A
公开(公告)日：1998-11-03
申请号：US943246
申请日：1992-09-10
申请人： Henryk Lubon , William N. Drohan , Lother Hennighausen
发明人： Henryk Lubon , William N. Drohan , Lother Hennighausen
IPC分类号： A01K67/027 , A61K38/43 , A61P7/02 , C12N9/64 , C12N15/09 , C12N15/57 , C12N15/85 , C12N5/00 , C12N15/00 , C12P21/04 , C12P21/06
CPC分类号： C12N15/8509 , A01K67/0278 , C12N9/6464 , C12Y304/21069 , A01K2207/15 , A01K2217/00 , A01K2217/05 , A01K2227/105 , A01K2227/108 , A01K2267/01 , C07K2319/02
摘要： Heterologous polypeptides are produced in the milk of transgenic non-human mammals by the expression of a stably integrated DNA sequence containing the long whey acidic protein promoter operably linked to a DNA sequence encoding a heterologous polypeptide and a signal sequence. The transgenic non-human mammals of the present invention are produced by introducing this DNA sequence such that the DNA sequence is stably integrated into the DNA of germ line cells of the mature mammal and inherited in normal Mendelian fashion. A representative heterologous polypeptide is protein C.
摘要翻译：通过表达含有可操作地连接到编码异源多肽和信号序列的DNA序列的长乳清酸性蛋白启动子的稳定整合的DNA序列，在转基因非人哺乳动物的乳中产生异源多肽。本发明的转基因非人哺乳动物是通过引入该DNA序列产生的，使得DNA序列稳定地整合到成熟哺乳动物的种系细胞的DNA中并以正常的孟德尔方式遗传。代表性的异源多肽是蛋白质C.

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