IP Library Granted Patent US 9,494,595
Granted Patent B2
US 9,494,595 · App. 14/363,201 · Granted Nov 15, 2016

Method for detecting protein-protein interaction

Inventors: Taku Watanabe (Ina, JP); Tatsuya Seki (Ina, JP); Aki Fujioka (Ina, JP)
Assignee: MEDICAL & BIOLOGICAL LABORATORIES CO., LTD.
G01N33/6845C12N15/85G01N33/542G01N33/582G01N2500/02
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Quick Facts
Patent No.
US 9,494,595
App. No.
14/363,201
Granted
Nov 15, 2016
Kind
B2
Abstract

A method for detecting an interaction between a first protein and a second protein comprises the steps of: expressing in a cell a first fusion protein comprising the first protein and an association-inducing protein, and a second fusion protein comprising the second protein and a fluorescent protein having a multimerization ability; detecting a fluorescent focus formed by an association between the first fusion protein and the second fusion protein in the cell; and determining an interaction between the first protein and the second protein according to the detection of the fluorescent focus.

Claims (9)

1. A method for detecting an interaction between a first protein and a second protein, the method comprising the steps of:

expressing in a cell a first fusion protein comprising the first protein and an association-inducing protein, and a second fusion protein comprising the second protein and a fluorescent protein having a multimerization ability and able of emitting fluorescence;

determining that there exists an interaction between the first protein and the second protein by detecting a fluorescent focus, wherein

the association-inducing protein is at least one protein selected from the group consisting of PB1 domain of p62, a PB1 domain of TFG, a PB1 domain of PKCiota, a SAM domain of TEL, a SAM domain of DGK delta, and a SAM domain of Tankyrase-1;

the fluorescent protein having a multimerization ability is at least one fluorescent protein selected from the group consisting of a fluorescent protein capable of forming a homomultimer, monomeric Kusabira-Orange 2, monomeric Keima-Red, monomeric Midoriishi-Cyan1, monomeric Kusabira-Orange 1, and monomeric Kikume Green-Red1; and

the fluorescent focus has a fluorescence intensity in a region of 0.2 to 5 μm, the fluorescence intensity being higher than a fluorescence intensity of the fluorescent protein which is present in a dispersed state in the cell, and wherein the association inducing protein is able to form a fluorescent focus when fused to the fluorescent protein having an multimerization ability.

2. The method of claim 1 , further detecting the presence of the interaction between the first protein and the second protein for a period of time wherein the absence of the focus indicates the ending of the interaction.

3. The method of claim 2 , further comprising adding a stimulus to the cell.

4. The method of claim 3 , wherein the interaction ends in response to a stimulus and further comprising measuring a time period from applying the stimulus to the cell to extincting the focus.

Assignments (2)
CHANGE OF ADDRESS Recorded Oct 28, 2022
From: MEDICAL & BIOLOGICAL LABORATORIES CO., LTD.
To: MEDICAL & BIOLOGICAL LABORATORIES CO., LTD.
Reel/Frame 061794/0822 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 29, 2014
From: WATANABE, TAKU; SEKI, TATSUYA; FUJIOKA, AKI
To: MEDICAL & BIOLOGICAL LABORATORIES CO., LTD.
Reel/Frame 033640/0359 →
Priority Claims (1)
JP 2011-266103 · Dec 5, 2011 · national
Continuity (1)
Related Publication 20140335539A1 · Nov 13, 2014