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    • 9. 发明授权
    • Mutant Aequorea victoria fluorescent proteins having increased cellular
fluorescence
    • 突变体Aequorea维多利亚荧光蛋白具有增加的细胞荧光
    • US6027881A
    • 2000-02-22
    • US646538
    • 1996-05-08
    • George N. PavlakisGeorge A. GaitanarisRoland H. StauberJohn N. Vournakis
    • George N. PavlakisGeorge A. GaitanarisRoland H. StauberJohn N. Vournakis
    • C12N15/09C07K14/435C12N5/10C12N15/12C12P21/02C12Q1/68C12R1/91C07H21/04C07K1/00C12N1/20
    • C07K14/43595C07K2319/00
    • The present invention is directed to mutants of the jellyfish Aequorea victoria green fluorescent protein (GFP) having at least 5 and preferably greater than 20 times the specific green fluorescence of the wild type protein. In other embodiments, the invention comprises mutant blue fluorescent proteins (BFPs) that emit an enhanced blue fluorescence. The invention also encompasses the expression of nucleic acids that encode a mutant GFP or BFP in a wide variety of engineered host cells, and the isolation of engineered proteins having increased fluorescent activity. The novel mutants of the present invention allow for a significantly more sensitive detection of fluorescence in engineered host cells than is possible with GFP or with its known mutants. Thus, the mutant fluorescent proteins provided herein can be used as sensitive reporter molecules to detect the cell and tissue-specific expression and subcellular compartmentalization of GFP or BFP mutants, or of chimeric proteins comprising GFP or BFP mutants fused to a regulatory sequence or to a second protein sequence.
    • 本发明涉及具有野生型蛋白质的特定绿色荧光的至少5倍,优选大于20倍的水母Aequorea维多利亚绿色荧光蛋白(GFP)的突变体。 在其它实施方案中,本发明包括发射增强的蓝色荧光的突变蓝色荧光蛋白(BFP)。 本发明还包括在多种工程化宿主细胞中编码突变型GFP或BFP的核酸的表达,以及具有增加的荧光活性的工程蛋白的分离。 与GFP或其已知突变体相比,本发明的新突变体允许在工程化宿主细胞中显着更灵敏地检测荧光。 因此,本文提供的突变荧光蛋白可用作敏感的报道分子,以检测GFP或BFP突变体或​​融合至调节序列的GFP或BFP突变体的嵌合蛋白的细胞和组织特异性表达和亚细胞区室化 第二个蛋白质序列。
    • 10. 发明授权
    • Mutant Aequorea victoria fluorescent proteins having increased cellular fluorescence
    • 突变体Aequorea维多利亚荧光蛋白具有增加的细胞荧光
    • US06265548B1
    • 2001-07-24
    • US09503222
    • 2000-02-11
    • George N. PavlakisGeorge A. GaitanarisRoland H. StauberJohn N. Vournakis
    • George N. PavlakisGeorge A. GaitanarisRoland H. StauberJohn N. Vournakis
    • C07K14435
    • C07K14/43595C07K2319/00
    • The present invention is directed to mutants of the jellyfish Aequorea victoria green fluorescent protein (GFP) having at least 5 and preferably greater than 20 times the specific green fluorescence of the wild type protein. In other embodiments, the invention comprises mutant blue fluorescent proteins (BFPs) that emit an enhanced blue fluorescence. The invention also encompasses the expression of nucleic acids that encode a mutant GFP or BFP in a wide variety of engineered host cells, and the isolation of engineered proteins having increased fluorescent activity. The novel mutants of the present invention allow for a significantly more sensitive detection of fluorescence in engineered host cells than is possible with GFP or with its known mutants. Thus, the mutant fluorescent proteins provided herein can be used as sensitive reporter molecules to detect the cell and tissue-specific expression and subcellular compartmentalization of GFP or BFP mutants, or of chimeric proteins comprising GFP or BFP mutants fused to a regulatory sequence or to a second protein sequence.
    • 本发明涉及具有野生型蛋白质的特定绿色荧光的至少5倍,优选大于20倍的水母Aequorea维多利亚绿色荧光蛋白(GFP)的突变体。 在其它实施方案中,本发明包括发射增强的蓝色荧光的突变蓝色荧光蛋白(BFP)。 本发明还包括在多种工程化宿主细胞中编码突变型GFP或BFP的核酸的表达,以及具有增加的荧光活性的工程蛋白的分离。 与GFP或其已知突变体相比,本发明的新突变体允许在工程化宿主细胞中显着更灵敏地检测荧光。 因此,本文提供的突变荧光蛋白可用作敏感的报道分子,以检测GFP或BFP突变体或​​融合至调节序列的GFP或BFP突变体的嵌合蛋白的细胞和组织特异性表达和亚细胞区室化 第二个蛋白质序列。