公开(公告)号：US06638732B1

公开(公告)日：2003-10-28

申请号：US09472065

申请日：1999-12-23

申请人： Krista Evans

发明人： Krista Evans

IPC分类号： C12P2106

CPC分类号： C07K14/43595

摘要： The present invention provides mutants of the Green Fluorescent Protein (GFP) of Aequorea victoria. Specifically provided by the present invention are nucleic acid molecules encoding mutant GFPs, the mutant GFPs encoded by these nucleic acid molecules, vectors and host cells comprising these nucleic acid molecules, and kits comprising one or more of the above as components. The invention also provides methods for producing these mutant GFPs. The fluorescence of these mutants is observable using fluorescein optics, making the mutant proteins of the present invention available for use in techniques such as fluorescence microscopy and flow cytometry using standard FITC filter sets. In addition, certain of these mutant proteins fluoresce when illuminated by white light, particularly when expressed at high levels in prokaryotic or eukaryotic host cells or when present in solution or in purified form at high concentrations. The mutant GFP sequences and peptides of the present invention are useful in the detection of transfection, in fluorescent labeling of proteins, in construction of fusion proteins allowing examination of intracellular protein expression, biochemistry and trafficking, and in other applications requiring the use of reporter genes.

摘要翻译： 本发明提供维他命A的绿色荧光蛋白（GFP）的突变体。 由本发明具体提供的是编码突变型GFP的核酸分子，由这些核酸分子编码的突变型GFP，包含这些核酸分子的载体和宿主细胞，以及包含上述一种或多种作为组分的试剂盒。 本发明还提供了用于产生这些突变GFP的方法。 这些突变体的荧光可以使用荧光素光学元件观察到，使得本发明的突变蛋白质可用于使用标准FITC过滤器组的技术如荧光显微镜和流式细胞术。 此外，当用白光照射时，这些突变体蛋白中的某些荧光发生荧光，特别是当在原核或真核宿主细胞中以高水平表达时，或当以高浓度存在于溶液或纯化形式时。 本发明的突变型GFP序列和肽可用于检测蛋白质的荧光标记中的转染，构建能够检测细胞内蛋白质表达，生物化学和运输的融合蛋白以及需要使用报道基因的其它应用 。

公开(公告)号：US20060229246A1

公开(公告)日：2006-10-12

申请号：US11333982

申请日：2006-01-17

申请人： Pamela Hawley-Nelson ,  Jianqing Lan ,  PoJen Shih ,  Joel Jessee ,  Keivn Schifferli ,  Gulilat Gebeyehu ,  Valentina Ciccarone ,  Krista Evans

发明人： Pamela Hawley-Nelson ,  Jianqing Lan ,  PoJen Shih ,  Joel Jessee ,  Keivn Schifferli ,  Gulilat Gebeyehu ,  Valentina Ciccarone ,  Krista Evans

IPC分类号： A61K38/17 ,  A61K48/00 ,  C12N15/88 ,  C12N15/86 ,  C12Q1/68

CPC分类号： B82Y5/00 ,  A61K9/1272 ,  A61K38/00 ,  A61K47/543 ,  A61K47/645 ,  A61K47/6911 ,  A61K48/00 ,  A61K48/0025 ,  A61K48/0041 ,  C07K1/1077 ,  C07K7/06 ,  C07K7/08 ,  C07K14/005 ,  C07K2319/09 ,  C12N15/87 ,  C12N15/88 ,  C12N2740/16322 ,  C12N2810/405 ,  C12N2810/60 ,  C12N2810/6018 ,  C12N2810/6054 ,  C12N2810/6081 ,  Y10T428/2984

摘要： The present invention provides compositions useful for transfecting eukaryotic cells comprising nucleic acid complexes with peptides, wherein the peptide is optionally covalently coupled to a nucleic acid-binding group, and cationic lipids or dendrimers as transfection agents. The invention also provides transfection compositions in which a peptide is covalently linked to the transfection agent (lipid, cationic lipid or dendrimer). Inclusion of peptides or modified-peptides in transfection compositions or covalent attachment of peptides to transfection agents results in enhanced transfection efficiency. Methods for the preparation of transfection compositions and methods of using these transfection compositions as intracellular delivery agents and extracellular targeting agents are also disclosed.

公开(公告)号：US20070021598A1

公开(公告)日：2007-01-25

申请号：US11399598

申请日：2006-04-07

申请人： Krista Evans

发明人： Krista Evans

IPC分类号： C07H21/04

CPC分类号： C07K14/43595

摘要： The present invention provides mutants of the Green Fluorescent Protein (GFP) of Aequorea victoria. Specifically provided by the present invention are nucleic acid molecules encoding mutant GFPs, the mutant GFPs encoded by these nucleic acid molecules, vectors and host cells comprising these nucleic acid molecules, and kits comprising one or more of the above as components. The invention also provides methods for producing these mutant GFPs. The fluorescence of these mutants is observable using fluorescein optics, making the mutant proteins of the present invention available for use in techniques such as fluorescence microscopy and flow cytometry using standard FITC filter sets. In addition, certain of these mutant proteins fluoresce when illuminated by white light, particularly when expressed at high levels in prokaryotic or eukaryotic host cells or when present in solution or in purified form at high concentrations. The mutant GFP sequences and peptides of the present invention are useful in the detection of transfection, in fluorescent labeling of proteins, in construction of fusion proteins allowing examination of intracellular protein expression, biochemistry and trafficking, and in other applications requiring the use of reporter genes.

摘要翻译： 本发明提供维他命A的绿色荧光蛋白（GFP）的突变体。 由本发明具体提供的是编码突变型GFP的核酸分子，由这些核酸分子编码的突变型GFP，包含这些核酸分子的载体和宿主细胞，以及包含上述一种或多种作为组分的试剂盒。 本发明还提供了用于产生这些突变GFP的方法。 这些突变体的荧光可以使用荧光素光学元件观察到，使得本发明的突变蛋白质可用于使用标准FITC过滤器组的技术如荧光显微镜和流式细胞术。 此外，当用白光照射时，这些突变体蛋白中的某些荧光发生荧光，特别是当在原核或真核宿主细胞中以高水平表达时，或当以高浓度存在于溶液或纯化形式时。 本发明的突变GFP序列和肽可用于检测蛋白质的荧光标记中的转染，构建能够检测细胞内蛋白质表达，生物化学和运输的融合蛋白以及需要使用报告基因的其它应用 。