公开(公告)号：US20050208624A1

公开(公告)日：2005-09-22

申请号：US11042158

申请日：2005-01-26

申请人： Atsushi Miyawaki ,  Takeharu Nagai

发明人： Atsushi Miyawaki ,  Takeharu Nagai

IPC分类号： G01N33/52 ,  C07K14/00 ,  C07K14/435 ,  C07K14/47 ,  C07K19/00 ,  C12N1/15 ,  C12N1/19 ,  C12N1/21 ,  C12N5/10 ,  C12N15/09 ,  C12Q1/02 ,  G01N21/78 ,  G01N33/533 ,  G01N33/58 ,  G01N33/84 ,  G01N33/20 ,  C07H21/04 ,  C12Q1/68

CPC分类号： G01N33/533 ,  C07K14/43595 ,  C07K14/4728 ,  C07K2319/00 ,  G01N33/582 ,  G01N33/84

摘要： It is an object of the present invention to provide a novel fluorescent protein having sensitivity to a calcium ion, in particular a fluorescent protein which can be used as an indicator for quantifying calcium ion concentration. The present invention provides a fluorescent protein having the following amino acid sequences (1) to (3) in order in the direction from the N-terminus to the C-terminus, wherein a fused fluorescent protein obtained by fusion of the fluorescent protein with a calcium binding protein and its target peptide can emit fluorescence which is dependent on Ca2+ ion level; (1) an amino acid sequence from the nth amino acid from the N-terminus to the C-terminus of a fluorescent protein selected from the group consisting of a green fluorescent protein or its mutant, a yellow fluorescent protein or its mutant, a cyan fluorescent protein or its mutant, a red fluorescent protein or its mutant, and a blue fluorescent protein or its mutant, provided that n represents an integer of 140 to 150; (2) a linker sequence of a sequence of 2 to 20 amino acids; and (3) an amino acid sequence from the 1st amino acid to the (n-1)th amino acid from the N-terminus of the fluorescent protein described in (1) above.

摘要翻译： 本发明的目的是提供一种对钙离子具有敏感性的新型荧光蛋白，特别是荧光蛋白，其可以用作定量钙离子浓度的指标。 本发明提供了从N末端到C末端的顺序依次具有以下氨基酸序列（1）〜（3）的荧光蛋白质，其中通过荧光蛋白质与 钙结合蛋白及其靶肽可以发射依赖于Ca 2+离子水平的荧光; （1）选自绿色荧光蛋白或其突变体，黄色的荧光蛋白的N末端至C末端的第n个氨基酸的氨基酸序列 荧光蛋白或其突变体，青色荧光蛋白或其突变体，红色荧光蛋白或其突变体，以及蓝色荧光蛋白或其突变体，条件是n表示140至150的整数; （2）2至20个氨基酸序列的接头序列; 和（3）从第（1）个氨基酸至（n-1）第（N）个氨基酸的氨基酸序列，从（ 1）以上。

公开(公告)号：US06627449B1

公开(公告)日：2003-09-30

申请号：US09704463

申请日：2000-10-31

申请人： Roger Y. Tsien ,  Atsushi Miyawaki ,  Juan Llopis

发明人： Roger Y. Tsien ,  Atsushi Miyawaki ,  Juan Llopis

IPC分类号： G01N3348

CPC分类号： C07K14/43595 ,  G01N33/84 ,  G01N2333/43595 ,  Y10S435/81

摘要： Disclosed are fluorescent protein sensors for measuring the pH of a sample, nucleic acids encoding them, and methods of use. The preferred fluorescent protein sensors are variants of the green fluorescent protein (GFP) from Aequorea victoria. Also disclosed are compositions and methods for measuring the pH of a specific region of a cell, such as the mitochondrial matrix or the Golgi lumen.

公开(公告)号：US06197928B1

公开(公告)日：2001-03-06

申请号：US08818252

申请日：1997-03-14

申请人： Roger Y. Tsien ,  Atsushi Miyawaki

发明人： Roger Y. Tsien ,  Atsushi Miyawaki

IPC分类号： C07K100

CPC分类号： C07K14/43595 ,  A01K2217/05 ,  C07K2319/00 ,  G01N33/542

摘要： Fluorescent indicators including a binding protein moiety, a donor fluorescent protein moiety, and an acceptor fluorescent protein moiety are described. The binding protein moiety has an analyte-binding region which binds an analyte and causes the indicator to change conformation upon exposure to the analyte. The donor moiety and the acceptor moiety change position relative to each other when the analyte binds to the analyte-binding region. The donor moiety and the acceptor moiety exhibit fluorescence resonance energy transfer when the donor moiety is excited and the distance between the donor moiety and the acceptor moiety is small. The indicators can be used to measure analyte concentrations in samples, such as calcium ion concentrations in cells.

摘要翻译： 描述了包括结合蛋白质部分，供体荧光蛋白部分和受体荧光蛋白部分的荧光指示剂。 结合蛋白部分具有结合分析物的分析物结合区域，并使得指示剂在暴露于分析物时改变构象。 当分析物结合到分析物结合区域时，供体部分和受体部分相对于彼此改变位置。 当供体部分被激发并且供体部分和受体部分之间的距离小时，供体部分和受体部分显示荧光共振能量转移。 指标可用于测量样品中的分析物浓度，如细胞中的钙离子浓度。

公开(公告)号：US5998204A

公开(公告)日：1999-12-07

申请号：US818253

申请日：1997-03-14

申请人： Roger Y. Tsien ,  Atsushi Miyawaki

发明人： Roger Y. Tsien ,  Atsushi Miyawaki

IPC分类号： C07K14/435 ,  G01N33/542 ,  C12N5/10 ,  C12N15/12 ,  C12N15/62 ,  C12N15/63

CPC分类号： G01N33/542 ,  C07K14/43595 ,  A01K2217/05 ,  C07K2319/00

摘要： Fluorescent indicators including a binding protein moiety, a donor fluorescent protein moiety, and an acceptor fluorescent protein moiety are described. The binding protein moiety has an analyte-binding region which binds an analyte and causes the indicator to change conformation upon exposure to the analyte. The donor moiety and the acceptor moiety change position relative to each other when the analyte binds to the analyte-binding region. The donor moiety and the acceptor moiety exhibit fluorescence resonance energy transfer when the donor moiety is excited and the distance between the donor moiety and the acceptor moiety is small. The indicators can be used to measure analyte concentrations in samples, such as calcium ion concentrations in cells.