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1. 发明申请

WO1997042320A1 MUTANT AEQUOREA VICTORIA FLUORESCENT PROTEINS HAVING INCREASED CELLULAR FLUORESCENCE 审中-公开
标题翻译：具有增加细胞荧光的突变AEOROREA VICTORIA荧光蛋白
公开(公告)号：WO1997042320A1
公开(公告)日：1997-11-13
申请号：PCT/US1997007625
申请日：1997-05-07
申请人： THE GOVERNMENT OF THE UNITED STATES OF AMERICA, represented by THE SECRETARY, DEPARTMENT OF HEALTH AND HUMAN SERVICES
发明人： THE GOVERNMENT OF THE UNITED STATES OF AMERICA, represented by THE SECRETARY, DEPARTMENT OF HEALTH AND HUMAN SERVICES , PAVLAKIS, George, N. , GAITANARIS, George, A. , STAUBER, Roland, H. , VOURNAKIS, John, N.
IPC分类号： C12N15/12
CPC分类号： C07K14/43595 , C07K2319/00
摘要： 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.
摘要翻译：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，以及分离具有增加的荧光活性的工程蛋白。与GFP或其已知突变体相比，本发明的新型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。

2. 发明申请

WO1984002534A1 HUMAN GROWTH HORMONE PRODUCED BY RECOMBINANT DNA IN MOUSE CELLS 审中-公开
标题翻译：人类细胞中重组DNA产生的人生长激素
公开(公告)号：WO1984002534A1
公开(公告)日：1984-07-05
申请号：PCT/US1983002016
申请日：1983-12-22
申请人： UNITED STATES OF AMERICA represented by THE ...
发明人： UNITED STATES OF AMERICA represented by THE ... , HAMER, Dean, H. , PAVLAKIS, George, N.
IPC分类号： C12P21/00
CPC分类号： C12N15/86 , C07K14/61 , C12N2710/20043 , C12N2830/002 , C12N2830/85
摘要： A recombinant DNA composed of (1) bovine papilloma virus, (2) the promoter region of the mouse metallothionein I gene and (3) human growth hormone structural sequences ligated to the metallothionein promoter was constructed. The recombinant was stably maintained as an episome and directed production of human growth hormone when introduced into cultured mammalian cells. Not only was the yield unexpectedly high, purification was vastly simplified because the growth hormone was secreted into the tissue culture medium. The process was suitable for spinner culture. Additionally, recombinant DNA molecules composed of (1) bovine papilloma virus and (2) the whole metallothionein I gene were utilized to render mouse cells resistant to toxic concentrations of cadmium. This combination was utilized as a selective marker to cotransfer other, non-selectable genes (such as human growth hormone) into mammalian cells.
摘要翻译：构建由（1）牛乳头瘤病毒，（2）小鼠金属硫蛋白I基因的启动子区和（3）连接到金属硫蛋白启动子的人生长激素结构序列组成的重组DNA。当引入培养的哺乳动物细胞中时，重组体被稳定地保持为附加体并且指导人生长激素的产生。不仅产量意外高，纯化被大大简化，因为生长激素分泌到组织培养基中。该过程适合旋转培养。此外，由（1）牛乳头瘤病毒和（2）整个金属硫蛋白I基因组成的重组DNA分子被用于使小鼠细胞对毒性浓度的镉具有抗性。该组合被用作选择性标记以将其他非选择性基因（例如人生长激素）转运到哺乳动物细胞中。

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