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1. 发明授权

US06593463B1 Modified MSP-1 nucleic acid sequences and methods for increasing MRNA levels and protein expression in cell systems 失效
标题翻译：改进的MSP-1核酸序列和用于增加细胞系统中MRNA水平和蛋白质表达的方法
公开(公告)号：US06593463B1
公开(公告)日：2003-07-15
申请号：US09175684
申请日：1998-10-20
申请人： Li How Chen , Harry Meade
发明人： Li How Chen , Harry Meade
IPC分类号： C07H2104
CPC分类号： C12N15/8509 , A01K67/0275 , A01K2217/05 , A01K2227/10 , A01K2227/105 , A01K2267/01 , A61K39/00 , A61K48/00 , A61K2039/51 , A61K2039/53 , C07K14/445 , C07K14/4732 , C07K2319/00 , C07K2319/02 , C12N15/67 , C12N2830/008 , Y02A50/412
摘要： The invention provides modified recombinant nucleic acid sequences (preferably DNA) and methods for increasing the mRNA levels and protein expression of malarial surface protein MSP-1 which is known to be difficult to express in cell culture systems, mammalian cell culture systems, or in transgenic animals. The preferred protein candidates for expression using the recombinant techniques of the invention are MSP-1 proteins expressed from DNA coding sequences comprising reduced overall AT content or AT rich regions and/or mRNA instability motifs and/or rare codons relative to the native MSP-1 gene.
摘要翻译：本发明提供修饰的重组核酸序列（优选DNA）和用于增加已知在细胞培养系统，哺乳动物细胞培养系统或转基因中已知难以表达的疟原虫表面蛋白质MSP-1的mRNA水平和蛋白质表达的方法动物。使用本发明的重组技术的用于表达的优选蛋白质候选物是由相对于天然MSP-1的包含减少的总体AT含量或AT富含区域和/或mRNA不稳定性基序和/或稀有密码子的DNA编码序列表达的MSP-1蛋白质基因。

2. 发明授权

US07501553B2 Non-human transgenic mammal comprising a modified MSP-1 nucleic acid 失效
标题翻译：包含修饰的MSP-1核酸的非人转基因哺乳动物
公开(公告)号：US07501553B2
公开(公告)日：2009-03-10
申请号：US10082018
申请日：2002-02-20
申请人： Li-How Chen , Harry M. Meade
发明人： Li-How Chen , Harry M. Meade
IPC分类号： C12P21/00 , A01K67/00
CPC分类号： C12N15/8509 , A01K67/0275 , A01K2217/05 , A01K2227/10 , A01K2227/105 , A01K2267/01 , A61K39/00 , A61K48/00 , A61K2039/51 , A61K2039/53 , C07K14/445 , C07K14/4732 , C07K2319/00 , C07K2319/02 , C12N15/67 , C12N2830/008 , Y02A50/412
摘要： The invention provides modified recombinant nucleic acid sequences (preferably DNA) and methods for increasing the mRNA levels and protein expression of malarial surface protein MSP-1 which is known to be difficult to express in cell culture systems, mammalian cell culture systems, or in transgenic animals. The preferred protein candidates for expression using the recombinant techniques of the invention are MSP-1 proteins expressed from DNA coding sequences comprising reduced overall AT content or AT rich regions and/or mRNA instability motifs and/or rare codons relative to the native MSP-1 gene.
摘要翻译：本发明提供修饰的重组核酸序列（优选DNA）和用于增加已知在细胞培养系统，哺乳动物细胞培养系统或转基因中已知难以表达的疟原虫表面蛋白质MSP-1的mRNA水平和蛋白质表达的方法动物。使用本发明的重组技术的用于表达的优选蛋白质候选物是由相对于天然MSP-1的包含减少的总体AT含量或AT富含区域和/或mRNA不稳定性基序和/或稀有密码子的DNA编码序列表达的MSP-1蛋白质基因。

3. 发明授权

US07354594B2 Merozoite surface protein 1 lacking glycosylation sites 失效
标题翻译：裂殖子表面蛋白1缺乏糖基化位点
公开(公告)号：US07354594B2
公开(公告)日：2008-04-08
申请号：US11140676
申请日：2005-05-27
申请人： Li How Chen , Harry M. Meade
发明人： Li How Chen , Harry M. Meade
IPC分类号： A61K39/015 , A61K39/02 , C12P21/00 , A01K67/027
CPC分类号： C12N15/8509 , A01K67/0275 , A01K2217/05 , A01K2227/10 , A01K2227/105 , A01K2267/01 , A61K39/00 , A61K48/00 , A61K2039/51 , A61K2039/53 , C07K14/445 , C07K14/4732 , C07K2319/00 , C07K2319/02 , C12N15/67 , C12N2830/008 , Y02A50/412
摘要： The invention provides modified recombinant nucleic acid sequences (preferably DNA) and methods for increasing the mRNA levels and protein expression of malarial surface protein MSP-1 which is known to be difficult to express in cell culture systems, mammalian cell culture systems, or in transgenic animals. The preferred protein candidates for expression using the recombinant techniques of the invention are MSP-1 proteins expressed from DNA coding sequences comprising reduced overall AT content or AT rich regions and/or mRNA instability motifs and/or rare codons relative to the native MSP-1 gene.
摘要翻译：本发明提供修饰的重组核酸序列（优选DNA）和用于增加已知在细胞培养系统，哺乳动物细胞培养系统或转基因中已知难以表达的疟原虫表面蛋白质MSP-1的mRNA水平和蛋白质表达的方法动物。使用本发明的重组技术的用于表达的优选蛋白质候选物是由相对于天然MSP-1的包含减少的总体AT含量或AT富含区域和/或mRNA不稳定性基序和/或稀有密码子的DNA编码序列表达的MSP-1蛋白质基因。

4. 发明授权

US07271312B1 Modified nucleic acid sequences and methods for increasing mRNA levels and protein expression in cell systems 有权
公开(公告)号：US07271312B1
公开(公告)日：2007-09-18
申请号：US09175683
申请日：1998-10-20
申请人： Li How Chen , Harry Meade
发明人： Li How Chen , Harry Meade
IPC分类号： C12P21/00 , A01K61/00
摘要： The invention provides modified recombinant nucleic acid sequences (preferably DNA) and methods for increasing the mRNA levels and protein expression of proteins which are known to be, or are likely to be, difficult to express in cell culture systems, mammalian cell culture systems, or in transgenic animals. The preferred “difficult” protein candidates for expression using the recombinant techniques of the invention are those proteins derived from heterologous cells preferably those of lower organisms such as parasites, bacteria, and virus, having DNA coding sequences comprising high overall AT content or AT rich regions and/or mRNA instability motifs and/or rare codons relative to the recombinant expression system to be used.

5. 发明申请

US20050060766A1 Expression of dominant negative transmembrane receptors in the milk of transgenic animals 审中-公开
标题翻译：转基因动物乳中显性负跨膜受体的表达
公开(公告)号：US20050060766A1
公开(公告)日：2005-03-17
申请号：US10941486
申请日：2004-09-15
申请人： Li-How Chen
发明人： Li-How Chen
IPC分类号： A01K67/027 , A61K20060101 , C07K14/715 , C07K14/81 , C12N15/85 , C12N15/877
CPC分类号： A01K67/0278 , A01K2207/15 , A01K2217/00 , A01K2217/05 , A01K2227/10 , A01K2227/102 , A01K2227/105 , A01K2267/01 , C07K14/7155 , C07K14/8128 , C12N15/8509 , C12N15/8772 , C12N2830/008
摘要： The present invention provides data to demonstrates the transgenic mammal production of membrane spanning receptor proteins in the milk of transgenic animals, offering a method of production of these proteins and dominant negative versions thereof for use as therapeutic molecules.
摘要翻译：本发明提供了数据，以证明在转基因动物的乳中的跨膜跨膜受体蛋白质的转基因哺乳动物产生，提供这些蛋白质的生产方法及其显性阴性形式用作治疗分子。

6. 发明授权

US06528699B1 Transgenically produced non-secreted proteins 失效
标题翻译：转基因产生的非分泌蛋白
公开(公告)号：US06528699B1
公开(公告)日：2003-03-04
申请号：US09028551
申请日：1998-02-24
申请人： Harry Meade , Li-How Chen , Paul DiTullio
发明人： Harry Meade , Li-How Chen , Paul DiTullio
IPC分类号： C12P2100
CPC分类号： C12N15/8509 , A01K67/0275 , A01K67/0278 , A01K2207/15 , A01K2217/00 , A01K2217/05 , A01K2227/102 , A01K2227/105 , A01K2267/01 , A61K38/00 , C07K14/4713 , C07K2319/036 , C12N9/88 , C12N2830/008
摘要： The invention provides a method of making and secreting a non-secreted protein. The method includes expressing the protein from a nucleic acid construct which includes: (a) a mammary epithelial specific promoter; (b) a milk protein specific signal sequence which can direct the secretion of a protein; (c) optionally, a sequence which encodes a sufficient portion of the amino terminal coding region of a secreted protein to allow secretion in the milk of a transgenic mammal, of the non-secreted protein; and (d) a sequence which encodes a non-secreted protein, wherein elements (a), (b), optionally (c), and (d) are preferably operatively linked in the order recited. Both glutamic acid decarboxylase (GAD) and myelin basic protein (MBP), which are cytoplasmic proteins, have been produced by the methods of the present invention. The invention also provides methods for treating diabetes and multiple sclerosis using proteins produced by the methods of the present invention.
摘要翻译：本发明提供了制备和分泌非分泌蛋白质的方法。该方法包括从核酸构建体表达蛋白质，其包括：（a）乳腺上皮特异性启动子; （b）可以指导蛋白质分泌的乳蛋白特异性信号序列; （c）任选地，编码分泌蛋白质的氨基末端编码区的足够部分以允许转基因哺乳动物的乳分泌的非分泌蛋白质的序列; 和（d）编码非分泌蛋白的序列，其中元件（a），（b），任选地（c）和（d）优选按所述顺序可操作地连接。通过本发明的方法制备了作为细胞质蛋白质的谷氨酸脱羧酶（GAD）和髓鞘碱性蛋白（MBP）。本发明还提供了使用通过本发明的方法产生的蛋白质来治疗糖尿病和多发性硬化的方法。

7. 发明申请

US20060191025A1 Method for the rapid selection of homozygous primary cell lines for the production of transgenic animals by somatic cell nuclear transfer 审中-公开
标题翻译：用于通过体细胞核转移快速选择用于产生转基因动物的纯合原代细胞系的方法
公开(公告)号：US20060191025A1
公开(公告)日：2006-08-24
申请号：US11408660
申请日：2006-04-21
申请人： Yann Echelard , Li-How Chen
发明人： Yann Echelard , Li-How Chen
IPC分类号： A01K67/027
CPC分类号： C12N15/8771 , A01K67/0273 , A01K2217/05 , A01K2227/10 , A01K2227/102 , A01K2267/01 , A01K2267/02 , C12N15/8509 , C12N15/877 , C12N15/8772
摘要： The present invention provides for the production of homozygous primary cells that carry a specific transgenic integration of interest on both chromosomes by bypassing breeding. These cell lines can they be used for the accelerated production of homozygous transgenic animals by somatic cell nuclear transfer. The invention is thus useful in the production of transgenic ungulate animals capable of producing desired biopharmaceuticals in their milk at higher yield than a comparable heterzygote. By combining the selection techniques of the current invention with somatic cell nuclear transfer it can be applied to large animals, where there is a strong need to shorten the time to homozygosity.
摘要翻译：本发明提供通过绕过育种在两条染色体上携带感兴趣的特定转基因整合的纯合原代细胞的生产。这些细胞系可以用于通过体细胞核转移加速纯合子转基因动物的产生。因此，本发明可用于生产能够以比同类杂合子更高的产量在其乳中产生所需生物药物的转基因有蹄类动物。通过将本发明的选择技术与体细胞核转移相结合，可以应用于大型动物，其中强烈需要缩短纯合子的时间。

8. 发明申请

US20060179493A1 Transgenically produced non-secreted proteins 审中-公开
公开(公告)号：US20060179493A1
公开(公告)日：2006-08-10
申请号：US11408119
申请日：2006-04-20
申请人： Harry Meade , Li-How Chen , Paul DiTullio
发明人： Harry Meade , Li-How Chen , Paul DiTullio
IPC分类号： A01K67/027
CPC分类号： C12N15/8509 , A01K67/0275 , A01K67/0278 , A01K2207/15 , A01K2217/00 , A01K2217/05 , A01K2227/102 , A01K2227/105 , A01K2267/01 , A61K38/00 , C07K14/4713 , C07K2319/036 , C12N9/88 , C12N2830/008
摘要： The invention provides a method of making and secreting a non-secreted protein. The method includes expressing the protein from a nucleic acid construct which includes: (a) a mammary epithelial specific promoter; (b) a milk protein specific signal sequence which can direct the secretion of a protein; (c) optionally, a sequence which encodes a sufficient portion of the amino terminal coding region of a secreted protein to allow secretion in the milk of a transgenic mammal, of the non-secreted protein; and (d) a sequence which encodes a non-secreted protein, wherein elements (a), (b), optionally (c), and (d) are preferably operatively linked in the order recited. Both glutamic acid decarboxylase (GAD) and myelin basic protein (MBP), which are cytoplasmic proteins, have been produced by the methods of the present invention. The invention also provides methods for treating diabetes and multiple sclerosis using proteins produced by the methods of the present invention.

9. 发明申请

US20060123500A1 Methods of prescreening cells for nuclear transfer procedures 审中-公开
标题翻译：预处理细胞核转移程序的方法
公开(公告)号：US20060123500A1
公开(公告)日：2006-06-08
申请号：US11081945
申请日：2005-03-16
申请人： Yann Echelard , Li-How Chen
发明人： Yann Echelard , Li-How Chen
IPC分类号： A01K67/027 , C12N5/06 , C12N15/87
CPC分类号： A01K67/0273 , A01K2227/102 , C12N2517/04
摘要： The present invention provides for the production of transgenic animals through pre-screening methods designed to improve the efficiency of nuclear transfer and consequentially the production characteristics of transgenic mammals relative to proteins of interest. The invention is thus useful in the production of transgenic ungulate animals capable of producing desired biopharmaceuticals in their milk at higher yield than a comparable heterzygote or producing animals with better physiological attributes.
摘要翻译：本发明提供了通过预筛选方法生产转基因动物，该方法旨在提高核转移的效率，从而相对于感兴趣的蛋白质的转基因哺乳动物的生产特性。因此，本发明可用于生产能够以比可比较的杂合子更高的产量在其乳中产生所需生物药物或生产具有更好的生理特性的动物的转基因有蹄类动物。

10. 发明授权

US07632980B1 Modified nucleic acid sequences and methods for increasing mRNA levels and protein expression in cell systems 失效
标题翻译：用于增加细胞系统中mRNA水平和蛋白质表达的修饰的核酸序列和方法
公开(公告)号：US07632980B1
公开(公告)日：2009-12-15
申请号：US09175683
申请日：1998-10-20
申请人： Li How Chen , Harry Meade
发明人： Li How Chen , Harry Meade
IPC分类号： C12P21/00 , A01K67/00
CPC分类号： C12N15/8509 , A01K67/0275 , A01K2217/05 , A01K2227/10 , A01K2227/105 , A01K2267/01 , A61K39/00 , A61K48/00 , A61K2039/51 , A61K2039/53 , C07K14/445 , C07K14/4732 , C07K2319/00 , C07K2319/02 , C12N15/67 , C12N2830/008 , Y02A50/412
摘要： The invention provides modified recombinant nucleic acid sequences (preferably DNA) and methods for increasing the mRNA levels and protein expression of proteins which are known to be, or are likely to be, difficult to express in cell culture systems, mammalian cell culture systems, or in transgenic animals. The preferred “difficult” protein candidates for expression using the recombinant techniques of the invention are those proteins derived from heterologous cells preferably those of lower organisms such as parasites, bacteria, and virus, having DNA coding sequences comprising high overall AT content or AT rich regions and/or mRNA instability motifs and/or rare codons relative to the recombinant expression system to be used.
摘要翻译：本发明提供了修饰的重组核酸序列（优选DNA）和用于增加已知在细胞培养系统，哺乳动物细胞培养系统中或可能难以在细胞培养系统中表达的蛋白质的mRNA水平和蛋白质表达的方法，或在转基因动物中。使用本发明的重组技术的优选的“困难”蛋白候选物是来自异源细胞的那些蛋白质，优选来自诸如寄生虫，细菌和病毒的较低生物体的蛋白质，其具有包含高总AT含量或AT丰富区域的DNA编码序列和/或mRNA不稳定性基序和/或稀有密码子相对于要使用的重组表达系统。

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