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1. 发明授权

US5391485A DNAs encoding analog GM-CSF molecules displaying resistance to proteases which cleave at adjacent dibasic residues 失效
标题翻译：编码模拟GM-CSF分子的DNA，其显示对相邻二碱基残基切割的蛋白酶的抗性
公开(公告)号：US5391485A
公开(公告)日：1995-02-21
申请号：US763130
申请日：1985-08-06
申请人： Michael C. Deeley , Virginia L. Price
发明人： Michael C. Deeley , Virginia L. Price
IPC分类号： C12N15/09 , A61K35/74 , A61K38/16 , C07K14/00 , C07K14/52 , C07K14/53 , C07K14/535 , C12N1/16 , C12N15/00 , C12N15/10 , C12N15/27 , C12P21/00 , C12P21/02 , C12R1/865 , C12N1/19 , C12N15/81
CPC分类号： C12N15/10 , C07K14/535 , Y10S930/145
摘要： Amplified expression of recombinant DNA products is achieved in hosts expressing proteases that cleave at multi-basic amino acid residues. To this end, cDNAs encoding granulocyte-macrophage colony stimulating factor (GM-CSF) are mutated such that one or both of the arginine residues at positions 23 and 24 of the protein product are replaced by non-basic amino acid residues. The GM-CSF analogs thus obtained maintain the activity of the wild-type protein.
摘要翻译：重组DNA产物的扩增表达在表达在多碱性氨基酸残基上切割的蛋白酶的宿主中实现。为此，编码粒细胞 - 巨噬细胞集落刺激因子（GM-CSF）的cDNA被突变，使得蛋白质产物的位置23和24处的精氨酸残基中的一个或两个被非碱性氨基酸残基替代。由此获得的GM-CSF类似物维持野生型蛋白质的活性。

2. 发明授权

US5393870A Analogs of human granulocyte-macrophage colony stimulating factor 失效
标题翻译：人粒细胞 - 巨噬细胞集落刺激因子的类似物
公开(公告)号：US5393870A
公开(公告)日：1995-02-28
申请号：US67934
申请日：1993-05-27
申请人： Michael C. Deeley , Virginia L. Price
发明人： Michael C. Deeley , Virginia L. Price
IPC分类号： C12N15/09 , A61K35/74 , A61K38/16 , C07K14/00 , C07K14/52 , C07K14/53 , C07K14/535 , C12N1/16 , C12N15/00 , C12N15/10 , C12N15/27 , C12P21/00 , C12P21/02 , C12R1/865 , C07K13/00
CPC分类号： C12N15/10 , C07K14/535 , Y10S930/145
摘要： Amplified expression of recombinant DNA products is achieved in hosts expressing proteases that cleave at multi-basic amino acid residues. To this end, cDNAs encoding granulocyte-macrophage colony stimulating factor (GM-CSF) are mutated such that one or both of the arginine residues at positions 23 and 24 of the protein product are deleted or replaced by non-basic amino acid residues. The GM-CSF analogs thus obtained maintain the activity of the wild-type protein.
摘要翻译：重组DNA产物的扩增表达在表达在多碱性氨基酸残基上切割的蛋白酶的宿主中实现。为此，编码粒细胞 - 巨噬细胞集落刺激因子（GM-CSF）的cDNA被突变，使得蛋白质产物的位置23和24处的精氨酸残基中的一个或两个被缺失或被非碱性氨基酸残基替代。由此获得的GM-CSF类似物维持野生型蛋白质的活性。

3. 发明授权

US5229496A Analogs of human granulocyte-macrophage colony stimulating factor 失效
标题翻译：人粒细胞 - 巨噬细胞集落刺激因子的类似物
公开(公告)号：US5229496A
公开(公告)日：1993-07-20
申请号：US254238
申请日：1988-10-06
申请人： Michael C. Deeley , Steven D. Gimpel , Virginia L. Price
发明人： Michael C. Deeley , Steven D. Gimpel , Virginia L. Price
IPC分类号： C12N15/09 , A61K35/74 , A61K38/16 , C07K14/00 , C07K14/52 , C07K14/53 , C07K14/535 , C12N1/16 , C12N15/00 , C12N15/10 , C12N15/27 , C12P21/00 , C12P21/02 , C12R1/865
CPC分类号： C12N15/10 , C07K14/535 , Y10S930/145
摘要： Amplified expression of recombinant DNA products is achieved in hosts expressing protease that cleave at multi-basic amino acid residues. To this end, wild-type genes encoding the desired protein products are mutated by substituting codons or eliminating codons encoding multi-basic amino acid residues while maintaining the activity of the expressed protein product. Mutation of the desired gene can be conveniently carried out by site-specific in vitro mutagenisis.
摘要翻译：重组DNA产物的扩增表达在表达在多碱性氨基酸残基处切割的蛋白酶的宿主中实现。为此，编码期望的蛋白质产物的野生型基因通过用密码子取代或消除编码多碱性氨基酸残基的密码子而突变，同时保持表达的蛋白质产物的活性。所需基因的突变可以通过位点特异性体外诱变方便地进行。

4. 发明申请

US20070254338A1 Method for making recombinant protein using complementation dependent DHFR mutants 审中-公开
标题翻译：使用互补依赖性DHFR突变体制备重组蛋白的方法
公开(公告)号：US20070254338A1
公开(公告)日：2007-11-01
申请号：US11789339
申请日：2007-04-24
申请人： R. Guy Caspary , Allison Bianchi , Pauline S. Smidt , Virginia L. Price , Martin J. Allen
发明人： R. Guy Caspary , Allison Bianchi , Pauline S. Smidt , Virginia L. Price , Martin J. Allen
IPC分类号： C12P21/06 , C12N9/06 , C07H21/04 , C12N15/74 , C12N1/21
CPC分类号： C12N9/003 , C12P21/02 , C12Y105/01003
摘要： The present invention relates to compositions and methods for making recombinant heteromeric proteins using a protein complementation assay employing complementation pairs of selectable markers.
摘要翻译：本发明涉及使用使用互补对选择标记的蛋白质互补测定法制备重组异聚蛋白质的组合物和方法。

5. 发明授权

US5616477A Fusion proteins comprising GM-CSF and antigens and their expression in yeast 失效
标题翻译：包含GM-CSF和抗原的融合蛋白及其在酵母中的表达
公开(公告)号：US5616477A
公开(公告)日：1997-04-01
申请号：US641704
申请日：1996-05-02
申请人： Virginia L. Price
发明人： Virginia L. Price
IPC分类号： A61K38/00 , C07K14/445 , C07K14/535 , C12N1/19 , C12N15/27 , C12N15/62
CPC分类号： C07K14/445 , C07K14/535 , A61K38/00 , C07K2319/00
摘要： Novel fusion proteins that enhance the immune response of an antigen are efficiently expressed and secreted by yeast host cells. The fusion proteins are recombinantly made by fusing the 3'-end of mature GM-CSF DNA sequence to the 5'-end of an antigen DNA sequence with or without a linker sequence. Methods of expression in yeast cells are provided.
摘要翻译：增强抗原免疫应答的新型融合蛋白由酵母宿主细胞有效表达和分泌。融合蛋白通过将成熟的GM-CSF DNA序列的3'末端与具有或不具有接头序列的抗原DNA序列的5'端融合来重组制备。提供酵母细胞中的表达方法。

6. 发明授权

US5538863A Expression system comprising mutant yeast strain and expression vector encoding synthetic signal peptide 失效
标题翻译：包含突变型酵母菌株和编码合成信号肽的表达载体的表达系统
公开(公告)号：US5538863A
公开(公告)日：1996-07-23
申请号：US86335
申请日：1993-07-01
申请人： Virginia L. Price
发明人： Virginia L. Price
IPC分类号： C07K14/535 , C07K14/54 , C12N15/62 , C12N15/81 , C12N1/19 , C12P21/02
CPC分类号： C07K14/5403 , C07K14/535 , C12N15/625 , C12N15/81 , C07K2319/036 , C07K2319/50 , C07K2319/75
摘要： A novel strain of Saccharomyces cerevisiae is useful as a host cell in the production of recombinant proteins. The novel S. cerevisiae cells transformed with a recombinant expression vector encoding a desired heterologous protein, preferably fused to a suitable N-terminal signal peptide, are cultivated under conditions that promote expression of the protein. Also provided are signal peptides derived by replacing the native signal peptidase cleavage site of a type I interleukin-1 receptor signal peptide with the tripeptide AlaXAla, wherein X represents an amino acid selected from Leu, Phe, and Gln. An expression system comprises a yeast host cell (preferably the novel S. cerevisiae strain) transformed with an expression vector comprising a promoter functional in yeast cells operably linked to DNA encoding the novel signal peptide, which is fused to the N-terminus of DNA encoding a desired heterologous protein.
摘要翻译：酿酒酵母的新型菌株可用作重组蛋白质生产中的宿主细胞。用促进蛋白质表达的条件培养用编码所需异源蛋白质的重组表达载体转化的新型酿酒酵母细胞，优选与合适的N-末端信号肽融合。还提供了通过用三肽AlaXAla替代I型白细胞介素-1受体信号肽的天然信号肽酶切割位点衍生的信号肽，其中X表示选自Leu，Phe和Gln的氨基酸。表达系统包括用表达载体转化的酵母宿主细胞（优选新型酿酒酵母菌株），该表达载体包含可操作地连接到编码新信号肽的DNA的酵母细胞中起作用的启动子，其与编码的DNA的N末端融合所需的异源蛋白。

7. 发明授权

US5122459A Gene encoding biologically active human interleukin 1 失效
标题翻译：基因编码生物活性人白细胞介素1
公开(公告)号：US5122459A
公开(公告)日：1992-06-16
申请号：US443399
申请日：1989-11-30
申请人： Paul J. Conlon, III , David j. Cosman , Kenneth H. Grabstein , Thomas P. Hopp , Shirley R. Kronheim , Alf D. Larsen , Carl J. March , Virginia L. Price , Douglas P. Cerretti
发明人： Paul J. Conlon, III , David j. Cosman , Kenneth H. Grabstein , Thomas P. Hopp , Shirley R. Kronheim , Alf D. Larsen , Carl J. March , Virginia L. Price , Douglas P. Cerretti
IPC分类号： C07K14/545 , C12N15/25 , C12N15/81
CPC分类号： C07K14/545 , C12N15/81 , Y10S435/942
摘要： Double-stranded cDNA is prepared from polyadenylated RNA extracted from activated human peripheral blood adherent mononuclear cells. The cDNA is inserted within a plasmid vector and then the recombinant plasmid employed to transform an appropriate host. Transformed hosts are identified and grouped into pools. Plasmid DNA prepared from these pools is hybridized with a labeled, synthetic oligonucleotide probe corresponding to a portion of the amino acid sequence of the interleukin 1 protein. Pools of host cells that provide a positive signal to the probe are identified, plated out and then employed in direct bacterial colony hybridization with the same probe, thereby to isolate the particular positive colony. Plasmid DNA is prepared from this colony and characterized by restriction enzyme mapping and sequencing by chain-termination method. The coding region for the IL-1 gene is inserted into a shuttle vector for amplification of the vector followed by expression of functional IL-1.
摘要翻译：双链cDNA由从活化的人外周血粘附单核细胞提取的聚腺苷酸化RNA制备。将cDNA插入质粒载体中，然后将重组质粒用于转化合适的宿主。已识别转化的主机并将其分组到池中。从这些池制备的质粒DNA与对应于白介素1蛋白的氨基酸序列的一部分的标记的合成的寡核苷酸探针杂交。确定提供探针的正信号的宿主细胞池，电镀，然后用于与相同探针的直接细菌菌落杂交，从而分离出特定的阳性菌落。从该菌落制备质粒DNA，并通过限制酶作图和通过链终止法进行测序。将用于IL-1基因的编码区插入穿梭载体中用于扩增载体，随后表达功能性IL-1。

8. 发明授权

US5405952A DNA sequence encoding nonglycosylated analogs of human colony stimulating factors 失效
标题翻译：编码人类集落刺激因子的非糖基化类似物的DNA序列
公开(公告)号：US5405952A
公开(公告)日：1995-04-11
申请号：US262385
申请日：1988-10-24
申请人： Michael Deeley , Virginia L. Price , David Urdal
发明人： Michael Deeley , Virginia L. Price , David Urdal
IPC分类号： C07K14/535 , C07H15/12 , C12N1/16 , C12N1/18 , C12N1/21 , C12N7/00 , C12N15/00 , C12P19/34 , C12P21/02 , C07K3/00
CPC分类号： C07K14/535 , Y10S530/82 , Y10S530/824
摘要： An analog human colony stimulating factor (hCSF) is disclosed, comprising a mutant amino acid sequence which is substantially homologous to the native sequence of an hCSF having at least one N-glycosylation site, wherein the mutant sequence comprises at least one amino acid substitution, deletion or insertion inactivating the N-glycosylation site.
摘要翻译：公开了类似人类集落刺激因子（hCSF），其包含与具有至少一个N-糖基化位点的hCSF的天然序列基本同源的突变氨基酸序列，其中突变序列包含至少一个氨基酸取代，缺失或插入使N-糖基化位点失活。

9. 发明授权

US5032676A Nonglycosylated analogs of human colony stimulating factors 失效
标题翻译：人类集落刺激因子的非糖基化类似物
公开(公告)号：US5032676A
公开(公告)日：1991-07-16
申请号：US918428
申请日：1986-10-14
申请人： Michael Deeley , Virginia L. Price , David Urdal
发明人： Michael Deeley , Virginia L. Price , David Urdal
IPC分类号： C07K14/535
CPC分类号： C07K14/535 , Y10S530/82 , Y10S530/824
摘要： An analog human colony stimulating factor (hCSF) is disclosed, comprising a mutant amino acid sequence which is substantially homologous to the native sequence of an hCSF having at least one N-glycosylation site, wherein the mutant sequence comprises at least one amino acid substitution, deletion or insertion inactivating the N-glycosylation site.
摘要翻译：公开了类似人类集落刺激因子（hCSF），其包含与具有至少一个N-糖基化位点的hCSF的天然序列基本同源的突变氨基酸序列，其中突变序列包含至少一个氨基酸取代，缺失或插入使N-糖基化位点失活。

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